U.S. patent application number 17/600305 was filed with the patent office on 2022-08-18 for ionic liquids for drug delivery.
This patent application is currently assigned to PRESIDENT AND FELLOWS OF HARVARD COLLEGE. The applicant listed for this patent is PRESIDENT AND FELLOWS OF HARVARD COLLEGE. Invention is credited to Samir MITRAGOTRI, Eden E.L. TANNER.
Application Number | 20220257767 17/600305 |
Document ID | / |
Family ID | 1000006347081 |
Filed Date | 2022-08-18 |
United States Patent
Application |
20220257767 |
Kind Code |
A1 |
MITRAGOTRI; Samir ; et
al. |
August 18, 2022 |
IONIC LIQUIDS FOR DRUG DELIVERY
Abstract
The technology described herein is directed to ionic liquids and
methods of drug delivery.
Inventors: |
MITRAGOTRI; Samir;
(LEXINGTON, MA) ; TANNER; Eden E.L.; (CAMBRIDGE,
MA) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
PRESIDENT AND FELLOWS OF HARVARD COLLEGE |
Cambridge |
MA |
US |
|
|
Assignee: |
PRESIDENT AND FELLOWS OF HARVARD
COLLEGE
Cambridge
MA
|
Family ID: |
1000006347081 |
Appl. No.: |
17/600305 |
Filed: |
March 26, 2020 |
PCT Filed: |
March 26, 2020 |
PCT NO: |
PCT/US2020/024866 |
371 Date: |
September 30, 2021 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
62828539 |
Apr 3, 2019 |
|
|
|
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 38/28 20130101;
A61K 31/519 20130101; A61K 47/12 20130101; A61K 9/4841 20130101;
A61K 47/186 20130101; A61K 38/26 20130101; A61K 9/08 20130101; A61K
9/5107 20130101; A61K 9/10 20130101 |
International
Class: |
A61K 47/18 20060101
A61K047/18; A61K 47/12 20060101 A61K047/12; A61K 38/26 20060101
A61K038/26; A61K 38/28 20060101 A61K038/28; A61K 31/519 20060101
A61K031/519; A61K 9/08 20060101 A61K009/08; A61K 9/10 20060101
A61K009/10; A61K 9/48 20060101 A61K009/48; A61K 9/51 20060101
A61K009/51 |
Claims
1. A method of administering at least one active compound, the
method comprising administering the active compound in combination
with at least one ionic liquid comprising: a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0; and a cation comprising a quaternary
ammonium.
2. A method of reducing weight/weight gain or treating obesity,
diabetes, ulcers, cancer, or fibrosis in a subject in need thereof,
the method comprising administering a composition comprising at
least one ionic liquid comprising: a hydrophobic anion comprising a
carboxylic acid having a pKa of at least 4.0 and a Log P of at
least 1.0; and a cation comprising a quaternary ammonium to the
subject.
3. The method of claim 2, wherein the composition does not comprise
a therapeutic agent other than the at least one ionic liquid.
4. The method of claim 2, wherein the composition further comprises
an active compound other than the at least one ionic liquid.
5. The method of any of the preceding claims, wherein the anion has
a pKa of at least 4.5.
6. The method of any of the preceding claims, wherein the anion has
a pKa of at least 5.0.
7. The method of any of the preceding claims, wherein the anion has
a pKa of at least 4.895.
8. The method of any of the preceding claims, wherein the anion has
a pKa of 4.5-5.5.
9. The method of any of the preceding claims, wherein the anion has
a pKa of 4.895-5.19.
10. The method of any of the preceding claims, wherein the anion
has a Log P of at least 2.0.
11. The method of any of the preceding claims, wherein the anion
has a Log P of at least 2.5.
12. The method of any of the preceding claims, wherein the anion
has a Log P of at least 2.75.
13. The method of any of the preceding claims, wherein the anion
has a Log P of at least 2.8.
14. The method of any of the preceding claims, wherein the anion
has a Log P of 2.5-3.5.
15. The method of any of the preceding claims, wherein the anion
has a Log P of 2.8-3.01.
16. The method of any of the preceding claims, wherein the anion
comprises a carbon chain of at least 8 carbons.
17. The method of any of the preceding claims, wherein the anion
comprises a carbon chain with an 8 carbon backbone.
18. The method of any of the preceding claims, wherein the anion is
geranic acid, octenoic acid, octanoic acid, or citronellic
acid.
19. The method of any of the preceding claims, wherein the anion is
octenoic acid, octanoic acid, or citronellic acid.
20. The method of any of the preceding claims, wherein the anion is
an alkene.
21. The method of any of the preceding claims, wherein the anion is
geranic acid, octanoic acid, or citronellic acid.
22. The method of any of the preceding claims, wherein the cation
has a molar mass equal to or greater than choline.
23. The method of any of the preceding claims, wherein the
quarternary ammonium has the structure of NR.sub.4.sup.+ and at
least one R group comprises a hydroxy group.
24. The method of any of the preceding claims, wherein the
quarternary ammonium has the structure of NR.sub.4.sup.+ and only
one R group comprises a hydroxy group.
25. The method of any of the preceding claims, wherein the cation
is C1, C6, or C7.
26. The method of any of the preceding claims, wherein the cation
is selected from choline, C1, C6, and C7 and the anion is
citronellic acid.
27. The method of any of the preceding claims, wherein the cation
is C1 and the anion is citronellic acid.
28. The method of any of the preceding claims, wherein the cation
is selected from C1, C6, and C7 and the anion is geranic acid.
29. The method of any of the preceding claims, wherein the ionic
liquid is choline: citronellic acid, C1: geranic acid, or C1:
citronellic acid.
30. The method of any of claims 1-16, wherein the cation is
selected from choline, C1, C6, and C7 and the anion is selected
from citronellic acid, octanoic acid, and octenoic acid.
31. The method of any of claims 1-16, wherein the cation is choline
and the anion is selected from citronellic acid, octanoic acid, and
octenoic acid.
32. The method of any of claims 1-16, wherein the ionic liquid is
choline: citronellic acid, choline: octanoic acid, or choline:
octenoic acid.
33. The method of any of the preceding claims, wherein the ionic
liquid is not CAGE.
34. The method of any of the preceding claims, wherein the ionic
liquid has less than 20 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
35. The method of any of the preceding claims, wherein the ionic
liquid has less than 10 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
36. The method of any of the preceding claims, wherein the ionic
liquid has less than 5 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
37. The method of any of the preceding claims, wherein the
administration is transdermal.
38. The method of any of the preceding claims, wherein the
administration is transdermal, to a mucus membrane, oral,
subcutaneous, intradermal, parenteral, intratumoral, or
intravenous.
39. The method of claim 26, wherein the mucus membrane is nasal,
oral, or vaginal.
40. The method of any of the preceding claims, wherein the
administration is oral.
41. The method of any of the preceding claims, wherein the ionic
liquid is at a concentration of at least 0.1% w/v.
42. The method of any of the preceding claims, wherein the ionic
liquid is at a concentration of from about 10 to about 70% w/v.
43. The method of any of the preceding claims, wherein the ionic
liquid is at a concentration of from about 30 to about 50% w/v.
44. The method of any of the preceding claims, wherein the ionic
liquid is at a concentration of from about 30 to about 40% w/v.
45. The method of any of the preceding claims, wherein the ionic
liquid comprises a ratio of cation to anion of from about 2:1 to
about 1:10.
46. The method of any of the preceding claims, wherein the ionic
liquid comprises a ratio of cation to anion of from about 1:1 to
about 1:4.
47. The method of any of the preceding claims, wherein the ionic
liquid comprises a ratio of cation to anion of about 1:2.
48. The method of any of the preceding claims, wherein the ionic
liquid has a cation:anion ratio of less than 1:1.
49. The method of any of the preceding claims, wherein the active
compound is hydrophobic.
50. The method of any of the preceding claims, wherein the active
compound is hydrophilic.
51. The method of any of the preceding claims, wherein the active
compound comprises a polypeptide.
52. The method of any of the preceding claims, wherein the active
compound has a molecular weight of greater than 450.
53. The method of any of the preceding claims, wherein the active
compound has a molecular weight of greater than 500.
54. The method of any of the preceding claims, wherein the active
compound comprises an antibody or antibody reagent.
55. The method of any of the preceding claims, wherein the active
compound comprises insulin, acarbose, ruxolitinib, or a GLP-1
polypeptide or mimetic or analog thereof.
56. The method of any of the preceding claims, wherein the
combination and/or composition is administered once.
57. The method of any of the preceding claims, wherein the
combination and/or composition is administered in multiple
doses.
58. The method of any of the preceding claims, wherein the active
compound and/or composition is provided at a dosage of 1-20
mg/kg.
59. The method of any of the preceding claims, wherein the active
compound and the ionic liquid are further in combination with at
least one non-ionic surfactant.
60. The method of any of the preceding claims, wherein the
combination and/or composition further comprises a further
pharmaceutically acceptable carrier.
61. The method of any of the preceding claims, wherein the
administration is oral and the combination and/or composition is
provided in a degradable capsule.
62. The method of any of the preceding claims, wherein the
combination is an admixture.
63. The method of any of the preceding claims, wherein the
combination and/or composition is provided in one or more
nanoparticles.
64. The method of any of the preceding claims, wherein the
combination is provided in the form of one or more nanoparticles
comprising the active compound, the nanoparticles in solution or
suspension in a composition comprising the ionic liquid.
65. A composition comprising at least one ionic liquid comprising:
a hydrophobic anion comprising a carboxylic acid having a pKa of at
least 4.0 and a Log P of at least 1.0; and a cation comprising a
quaternary ammonium.
66. The composition of claim 65, wherein the anion has a pKa of at
least 4.5.
67. The composition of claim 65, wherein the anion has a pKa of at
least 4.895.
68. The composition of claim 65, wherein the anion has a pKa of
4.5-5.5.
69. The composition of claim 65, wherein the anion has a pKa of
4.895-5.19.
70. The composition of any of claims 65-69, wherein the anion has a
pKa of at least 5.0.
71. The composition of any of claims 65-69, wherein the anion has a
Log P of at least 2.0.
72. The composition of any of claims 65-69, wherein the anion has a
Log P of at least 2.5.
73. The composition of any of claims 65-69, wherein the anion has a
Log P of at least 2.75.
74. The composition of any of claims 65-69, wherein the anion has a
Log P of at least 2.8.
75. The composition of any of claims 65-69, wherein the anion has a
Log P of 2.5-3.5.
76. The composition of any of claims 65-69, wherein the anion has a
Log P of 2.8-3.01.
77. The composition of any of claims 65-76, wherein the anion
comprises a carbon chain of at least 8 carbons.
78. The composition of any of claims 65-76, wherein the anion
comprises a carbon chain with an 8 carbon backbone.
79. The composition of any of claims 65-76, wherein the anion is
geranic acid, octenoic acid, octanoic acid, or citronellic
acid.
80. The composition of any of claims 65-76, wherein the anion is
octenoic acid, octanoic acid, or citronellic acid.
81. The composition of any of claims 65-80, wherein the anion is an
alkene.
82. The composition of any of claims 65-81, wherein the anion is
geranic acid, octanoic acid, or citronellic acid.
83. The composition of any of claims 65-82, wherein the cation has
a molar mass equal to or greater than choline.
84. The composition of any of claims 65-83, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and at least one R
group comprises a hydroxy group.
85. The composition of any of claims 65-84, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and only one R group
comprises a hydroxy group.
86. The composition of any of claims 65-85, wherein the cation is
C1, C6, or C7.
87. The composition of any of claims 65-86, wherein the cation is
selected from choline, C1, C6, and C7 and the anion is citronellic
acid.
88. The composition of any of claims 65-87, wherein the cation is
C1 and the anion is citronellic acid.
89. The composition of any of claims 65-88, wherein the cation is
selected from C1, C6, and C7 and the anion is geranic acid.
90. The composition of any of claims 65-89, wherein the ionic
liquid is choline: citronellic acid, C1: geranic acid, or C1:
citronellic acid.
91. The composition of any of claims 65-90, wherein the cation is
selected from choline, C1, C6, and C7 and the anion is selected
from citronellic acid, octanoic acid, and octenoic acid.
92. The composition of any of claims 65-91, wherein the cation is
choline and the anion is selected from citronellic acid, octanoic
acid, and octenoic acid.
93. The composition of any of claims 65-92, wherein the ionic
liquid is choline:citronellic acid, choline:octanoic acid, or
choline:octenoic acid.
94. The composition of any of claims 65-93, wherein the ionic
liquid is not CAGE.
95. The composition of any of claims 65-94, wherein the ionic
liquid comprises a ratio of cation to anion of from about 2:1 to
about 1:10.
96. The composition of any of claims 65-95, wherein the ionic
liquid comprises a ratio of cation to anion of from about 1:1 to
about 1:4.
97. The composition of any of claims 65-96, wherein the ionic
liquid comprises a ratio of cation to anion of about 1:2.
98. The composition of any of claims 65-97, wherein the ionic
liquid has a cation:anion ratio of less than 1:1.
99. The composition of any of claims 65-98, wherein the ionic
liquid has a cation:anion ratio with an excess of anion.
100. The composition of any of claims 65-99, wherein the ionic
liquid has less than 20 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
101. The composition of any of claims 65-100, wherein the ionic
liquid has less than 10 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
102. The composition of any of claims 65-101, wherein the ionic
liquid has less than 5 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY).
103. The composition of any of claims 65-102, further comprising at
least one active compound in combination with the at least one
ionic liquid.
104. The composition of claim 103, wherein the active compound is
hydrophobic.
105. The composition of any of claims 103-104, wherein the active
compound is hydrophilic.
106. The composition of any of claims 103-105, wherein the active
compound comprises a polypeptide.
107. The composition of any of claims 103-106, wherein the active
compound has a molecular weight of greater than 450.
108. The composition of any of claims 103-107, wherein the active
compound has a molecular weight of greater than 500.
109. The composition of any of claims 103-108, wherein the active
compound comprises an antibody or antibody reagent.
110. The composition of any of claims 103-109, wherein the active
compound comprises insulin, acarbose, ruxolitinib, or a GLP-1
polypeptide or mimetic or analog thereof.
111. The composition of any of claims 65-110, wherein the ionic
liquid is at a concentration of at least 0.1% w/v.
112. The composition of any of claims 65-111, wherein the ionic
liquid is at a concentration of from about 10 to about 70% w/v.
113. The composition of any of claims 65-112, wherein the ionic
liquid is at a concentration of from about 30 to about 50% w/v.
114. The composition of any of claims 65-113, wherein the ionic
liquid is at a concentration of from about 30 to about 40% w/v.
115. The composition of any of claims 65-114, wherein the
composition is formulated for transdermal administration.
116. The composition of any of claims 65-115, wherein the
composition is formulated for administration transdermally, to a
mucus membrane, orally, subcutaneously, intradermally,
parenterally, intratumorally, or intravenously.
117. The composition of claim 116, wherein the mucus membrane is
nasal, oral, or vaginal.
118. The composition of any of claims 65-117, wherein the
composition is formulated for oral administration.
119. The composition of any of claims 103-118, wherein the active
compound is provided at a dosage of 1-20 mg/kg.
120. The composition of any of claims 65-119, further comprising at
least one non-ionic surfactant.
121. The composition of any of claims 65-120, further comprising a
pharmaceutically acceptable carrier.
122. The composition of any of claims 65-121, wherein the
composition is provided in a degradable capsule.
123. The composition of any of claims 65-122, wherein the
composition is an admixture.
124. The composition of any of claims 65-123, wherein the
composition is provided in one or more nanoparticles.
125. The composition of any of claims 65-124, comprising one or
more nanoparticles comprising the active compound, the
nanoparticles in solution or suspension in a composition comprising
the ionic liquid.
126. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, the method comprising: a. selecting one of the two
ions of the ionic liquid; and b. selecting the other ion to
minimize inter-ionic interactions.
127. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, the method comprising: a. selecting the cation; and b.
selecting the anion to minimize inter-ionic interactions.
128. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, the method comprising: a. selecting the anion; and b.
selecting the cation to minimize inter-ionic interactions.
129. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, from a pool of candidate cations and a pool of
candidate anions, the method comprising: a. selecting one of the
two ions of the ionic liquid from the pool of candidate ions; and
b. selecting from the other pool of candidate ions the ion which
most minimizes inter-ionic interactions with the ion selected in
step a.
130. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, from a pool of candidate cations and a pool of
candidate anions, the method comprising: a. selecting the cation
from the pool of candidate cations; b. selecting from the pool of
candidate anions the anion which most minimizes inter-ionic
interactions with the cation selected in step a.
131. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, from a pool of candidate cations and a pool of
candidate anions, the method comprising: a. selecting the cation
from the pool of candidate anions; b. selecting from the pool of
candidate cations the anion which most minimizes inter-ionic
interactions with the anion selected in step a.
132. The method of any of claims 126-131, wherein the ionic liquid
is selected or designed for transdermal administration.
133. The method of any of claims 126-132, wherein the ionic liquid
is selected or designed for administration transdermally, to a
mucus membrane, orally, subcutaneously, intradermally,
parenterally, intratumorally, or intravenously.
134. The method of claim 133, wherein the mucus membrane is nasal,
oral, or vaginal.
135. The method of any of claims 126-134, wherein the ionic liquid
is selected or designed for oral administration.
136. The method of any of claims 126-135, wherein the ionic liquid
is selected or designed for delivery of an active compound.
137. The method of any of claims 126-136, wherein the cation
comprises, or selecting the cation comprises selection a cation
that comprises a quaternary ammonium; and the anion comprises or
selecting an anion comprises selecting a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0.
138. The method of any of claims 126-137, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
at least 4.5.
139. The method of any of claims 126-138, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
at least 4.895.
140. The method of any of claims 126-138, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
4.5-5.5.
141. The method of any of claims 126-140, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
4.895-5.19.
142. The method of any of claims 126-141, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
at least 5.0.
143. The method of any of claims 126-142, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of at least 2.0.
144. The method of any of claims 126-143, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of at least 2.5.
145. The method of any of claims 126-144, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of at least 2.75.
146. The method of any of claims 126-145, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of at least 2.8.
147. The method of any of claims 126-146, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of 2.5-3.5.
148. The method of any of claims 126-147, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a Log P
of 2.8-3.01.
149. The method of any of claims 126-148, wherein the anion
comprises, or selecting an anion comprises selecting an anion that
comprises, a carbon chain of at least 8 carbons.
150. The method of any of claims 126-149, wherein the anion
comprises, or selecting an anion comprises selecting an anion that
comprises, a carbon chain with an 8 carbon backbone.
151. The method of any of claims 126-150, wherein the anion is, or
selecting an anion comprises selecting an anion that is, geranic
acid, octenoic acid, octanoic acid, or citronellic acid.
152. The method of any of claims 126-151, wherein the anion is, or
selecting an anion comprises selecting an anion that is, octenoic
acid, octanoic acid, or citronellic acid.
153. The method of any of claims 126-152, wherein the anion is, or
selecting an anion comprises selecting an anion that is, an
alkene.
154. The method of any of claims 126-153, wherein the anion is, or
selecting an anion comprises selecting an anion that is, geranic
acid, octanoic acid, or citronellic acid.
155. The method of any of claims 126-154, wherein the cation has,
or selecting a cation comprises selecting a cation that has, a
molar mass equal to or greater than choline.
156. The method of any of claims 126-155, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and at least one R
group comprises a hydroxy group.
157. The method of any of claims 126-156, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and only one R group
comprises a hydroxy group.
158. The method of any of claims 126-157, wherein the cation is, or
selecting a cation comprises selecting a cation that is, C1, C6, or
C7.
159. The method of any of claims 126-158, wherein the cation is, or
selecting a cation comprises selecting a cation that is, selected
from choline, C1, C6, and C7 and the anion is citronellic acid.
160. The method of any of claims 126-159, wherein the cation is, or
selecting a cation comprises selecting a cation that is, selected
from C1, C6, and C7 and the anion is geranic acid.
161. The method of any of claims 126-160, wherein the cation is, or
selecting a cation comprises selecting a cation that is, choline,
C1, C6, or C7 and selecting the anion comprises selecting an anion
that is citronellic acid, octanoic acid, or octenoic acid.
162. The method of any of claims 126-161, wherein the cation is
choline and selecting the anion comprises selecting an anion that
is citronellic acid, octanoic acid, or octenoic acid.
163. The method of any of claims 126-162, wherein the ionic liquid
is not CAGE.
164. The method of any of claims 126-163, wherein the ionic liquid
comprises a ratio of cation to anion of from about 2:1 to about
1:10.
165. The method of any of claims 126-164, wherein the ionic liquid
comprises a ratio of cation to anion of from about 1:1 to about
1:4.
166. The method of any of claims 126-165, wherein the ionic liquid
comprises a ratio of cation to anion of about 1:2.
167. The method of any of claims 126-166, wherein the ionic liquid
has a cation:anion ratio of less than 1:1.
168. The method of any of claims 126-167, wherein the ionic liquid
has a cation:anion ratio with an excess of anion.
169. The method of any of claims 126-168, wherein minimizing
inter-ionic interaction comprises minimizing the number of cross
peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY).
170. The method of any of claims 126-169, wherein a cation and
anion minimize inter-ionic interaction if they have less than 20
cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY).
171. The method of any of claims 126-170, wherein a cation and
anion minimize inter-ionic interaction if they have less than 10
cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY).
172. The method of any of claims 126-171, wherein a cation and
anion minimize inter-ionic interaction if they have less than 5
cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY).
173. The method of any of claims 126-172, wherein the active
compound is hydrophobic.
174. The method of any of claims 126-173, wherein the active
compound is hydrophilic.
175. The method of any of claims 126-174, wherein the active
compound comprises a polypeptide.
176. The method of any of claims 126-175, wherein the active
compound has a molecular weight of greater than 450.
177. The method of any of claims 126-176, wherein the active
compound has a molecular weight of greater than 500.
178. The method of any of claims 126-177, wherein the active
compound comprises an antibody or antibody reagent.
179. The method of any of claims 126-178, wherein the active
compound comprises insulin, acarbose, ruxolitinib, or a GLP-1
polypeptide or mimetic or analog thereof.
180. The method of any of claims 126-179, wherein the ionic liquid
is at a concentration of at least 0.1% w/v.
181. The method of any of claims 126-180, wherein the ionic liquid
is at a concentration of from about 10 to about 70% w/v.
182. The method of any of claims 126-181, wherein the ionic liquid
is at a concentration of from about 30 to about 50% w/v.
183. The method of any of claims 126-182, wherein the ionic liquid
is at a concentration of from about 30 to about 40% w/v.
184. The method of any of claims 126-183, wherein the active
compound is provided at a dosage of 1-20 mg/kg.
185. The method of any of claims 126-184, wherein the ionic liquid
is designed or selected to be provided in a degradable capsule.
186. The method of any of claims 126-185, wherein the ionic liquid
and active compound are in admixture.
187. The method of any of claims 126-186, wherein the ionic liquid
and optionally the active compound are selected or designed to be
provided in one or more nanoparticles.
188. The method of any of claims 126-187, wherein the ionic liquid
is in solution or suspension in a composition with one or more
nanoparticles comprising the active compound.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims benefit under 35 U.S.C. .sctn.
119(e) of U.S. Provisional Application No. 62/828,539 filed Apr. 3,
2019, the contents of which are incorporated herein by reference in
their entirety.
SEQUENCE LISTING
[0002] The instant application contains a Sequence Listing which
has been submitted electronically in ASCII format and is hereby
incorporated by reference in its entirety. Said ASCII copy, created
on Mar. 24, 2020, is named 002806-094500WOPT_SL.txt and is 2,891
bytes in size.
TECHNICAL FIELD
[0003] The technology described herein relates to ionic liquids for
stabilization and delivery of active compounds.
BACKGROUND
[0004] The uptake of many active compounds, e.g., pharmaceutically
active compounds, can be improved by delivering the compounds in
solvents. However, such approaches are often unsuitable for in vivo
use because most such solvents demonstrate toxic side effects
and/or act as irritants to the point of delivery. These toxic and
irritant effects are severe enough to mitigate any increase in the
uptake or performance of the active compound.
SUMMARY
[0005] As demonstrated herein, the inventors have identified
characteristics of ionic liquids that provide surprising superior
active compound uptake kinetics. Accordingly, compositions and
methods relating to these ionic liquids (ILs) with unexpectedly
high efficacy are described herein.
[0006] In one aspect of any of the embodiments, described herein is
a method of administering at least one active compound, the method
comprising administering the active compound in combination with at
least one ionic liquid (IL) comprising: i) a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0; and ii) a cation comprising a quaternary
ammonium. In one aspect of any of the embodiments, described herein
is a combination of at least one active compound and at least one
ionic liquid comprising: i) a hydrophobic anion comprising a
carboxylic acid having a pKa of at least 4.0 and a Log P of at
least 1.0; and ii) a cation comprising a quaternary ammonium for
use in a method of drug delivery.
[0007] In one aspect of any of the embodiments, described herein is
a method of reducing weight/weight gain or treating obesity,
diabetes, ulcers, cancer, or fibrosis in a subject in need thereof,
the method comprising administering a composition comprising at
least one ionic liquid comprising: i) a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0; and ii) a cation comprising a quaternary
ammonium to the subject. In one aspect of any of the embodiments,
described herein is a composition comprising at least one ionic
liquid comprising: i) a hydrophobic anion comprising a carboxylic
acid having a pKa of at least 4.0 and a Log P of at least 1.0; and
ii) a cation comprising a quaternary ammonium for use in a method
of reducing weight/weight gain or treating obesity, diabetes,
ulcers, cancer, or fibrosis in a subject in need thereof. In some
embodiments, the subject is not administered, and/or the
composition does not comprise a therapeutic agent or active
compound other than the at least one ionic liquid. In some
embodiments, the subject is administered (e.g., in the same
formulation as the at least one IL or in a separate formulation),
and/or the composition further comprises a therapeutic agent or
active compound other than the at least one ionic liquid.
[0008] In one aspect, provided herein is a composition comprising
at least one ionic liquid comprising: i) a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0; and ii) a cation comprising a quaternary
ammonium. In some embodiments, the composition further comprises a
therapeutic agent or active compound in combination with the at
least one ionic liquid.
[0009] In some embodiments of any of the aspects, the anion has a
pKa of at least 4.5. In some embodiments of any of the aspects, the
anion has a pKa of at least 5.0. In some embodiments of any of the
aspects, the anion has a Log P of at least 2.0. In some embodiments
of any of the aspects, the anion has a Log P of at least 2.5. In
some embodiments of any of the aspects, the anion has a Log P of at
least 2.75.
[0010] In some embodiments of any of the aspects, the anion
comprises a carbon chain backbone of at least 8 carbons. In some
embodiments of any of the aspects, the anion is an alkene. In some
embodiments of any of the aspects, the anion is geranic acid,
octanoic acid, or citronellic acid.
[0011] In some embodiments of any of the aspects, the cation has a
molar mass equal to or greater than choline. In some embodiments of
any of the aspects, the cation has a molar mass greater than
choline. In some embodiments of any of the aspects, the quarternary
ammonium has the structure of NR.sub.4.sup.+ and at least one R
group comprises a hydroxy group. In some embodiments of any of the
aspects, the quarternary ammonium has the structure of
NR.sub.4.sup.+ and only one R group comprises a hydroxy group. In
some embodiments of any of the aspects, the cation is C1, C6, or
C7.
[0012] In some embodiments of any of the aspects, the cation is
selected from choline, C1, C6, and C7 and the anion is citronellic
acid. In some embodiments of any of the aspects, the cation is C1
and the anion is citronellic acid. In some embodiments of any of
the aspects, the cation is selected from C1, C6, and C7 and the
anion is geranic acid. In some embodiments of any of the aspects,
the ionic liquid is choline: citronellic acid, C1: geranic acid, or
C1: citronellic acid. In some embodiments of any of the aspects,
the ionic liquid is not CAGE.
[0013] In some embodiments of any of the aspects, the ionic liquid
has less than 20, less than 10, or less than 5 cross peaks as
measured by Nuclear Overhauser Effect SpectroscopY (NOESY).
[0014] In some embodiments of any of the aspects, the
administration is transdermal. In some embodiments of any of the
aspects, the administration is transdermal, to a mucus membrane,
oral, subcutaneous, intradermal, parenteral, intratumoral, or
intravenous. In some embodiments of any of the aspects, the
composition or combination is formulated for transdermal
administration. In some embodiments of any of the aspects, the
composition or combination is formulated for transdermal, to a
mucus membrane, oral, subcutaneous, intradermal, parenteral,
intratumoral, or intravenous administration. In some embodiments of
any of the aspects, the mucus membrane is nasal, oral, or
vaginal.
[0015] In some embodiments of any of the aspects, the ionic liquid
is at a concentration of at least 0.1% w/v. In some embodiments of
any of the aspects, the ionic liquid is at a concentration of from
about 10 to about 70% w/v. In some embodiments of any of the
aspects, the ionic liquid is at a concentration of from about 30 to
about 50% w/v. In some embodiments of any of the aspects, the ionic
liquid is at a concentration of from about 30 to about 40% w/v. In
some embodiments of any of the aspects, the ionic liquid comprises
a ratio of cation to anion of from about 2:1 to about 1:10. In some
embodiments of any of the aspects, the ionic liquid comprises a
ratio of cation to anion of from about 1:1 to about 1:4. In some
embodiments of any of the aspects, the ionic liquid comprises a
ratio of cation to anion of about 1:2. In some embodiments of any
of the aspects, the ionic liquid has a cation:anion ratio of less
than 1:1. In some embodiments of any of the aspects, the ionic
liquid has a cation:anion ratio comprising an excess of anion.
[0016] In some embodiments of any of the aspects, the active
compound is hydrophobic. In some embodiments of any of the aspects,
the active compound is hydrophilic. In some embodiments of any of
the aspects, the active compound comprises a polypeptide. In some
embodiments of any of the aspects, the active compound has a
molecular weight of greater than 450. In some embodiments of any of
the aspects, the active compound has a molecular weight of greater
than 500. In some embodiments of any of the aspects, the active
compound comprises an antibody or antibody reagent. In some
embodiments of any of the aspects, the active compound comprises
insulin, acarbose, ruxolitinib, or a GLP-1 polypeptide or mimetic
or analog thereof.
[0017] In some embodiments of any of the aspects, the combination
and/or composition is administered once. In some embodiments of any
of the aspects, the combination and/or composition is administered
in multiple doses. In some embodiments of any of the aspects, the
active compound and/or composition is provided at a dosage of 1-20
mg/kg.
[0018] In some embodiments of any of the aspects, the active
compound and the ionic liquid are further in combination with at
least one non-ionic surfactant. In some embodiments of any of the
aspects, the combination and/or composition further comprises a
further pharmaceutically acceptable carrier. In some embodiments of
any of the aspects, the administration is oral and the combination
and/or composition is provided in a degradable capsule. In some
embodiments of any of the aspects, the combination is an admixture.
In some embodiments of any of the aspects, the combination and/or
composition is provided in one or more nanoparticles. In some
embodiments of any of the aspects, the combination is provided in
the form of one or more nanoparticles comprising the active
compound, the nanoparticles in solution or suspension in a
composition comprising the ionic liquid.
BRIEF DESCRIPTION OF THE DRAWINGS
[0019] FIG. 1A depicts the chemical structures of Choline and
Geranic acid. FIG. 1B depicts CAGE prepared by salt metathesis of
choline bicarbonate and geranic acid at a molar ratio of 1:2.
[0020] FIG. 2 depicts the two drugs assessed in this study:
acarbose and ruxolitinib. Acarbose was used as model hydrophilic
drug (MW: 646, Log P: -6.8) and ruxolitinib was used as a model
hydrophobic drug (MW: 306 and Log P: 2.9).
[0021] FIG. 3 depicts the amount of acarbose or ruxolitinib
delivered into and across the skin after topical application from
various CAGE compositions. Both drugs were dissolved in CAGE at a
concentration of 1 mg/ml (N=3, error bars are SEM).
[0022] FIG. 4 depicts the anions used as alternatives to geranic
acid. The anions were selected to cover a range of molecular
weights, pKa and Log P values.
[0023] FIGS. 5A-5D depict the relationships between the overall
transport rank of the anionic alternatives and the (FIG. 5A)
molecular weight, (FIG. 5B) pKa, (FIG. 5C) Log P and (FIG. 5D) the
number of carbons in the anion.
[0024] FIGS. 6A-6B depict the NOESY spectra of (FIG. 6A) choline:
citronellic acid and (FIG. 6B) choline: glutaric acid. The circled
cross peaks represent parts of the molecule are within 5 nm of each
other. The number of cross peaks exhibit a significant difference
among the ILs studied.
[0025] FIG. 7 depicts the correlation between the transport ranking
and number of NOSEY cross correlation peaks. NOSEY data for
choline: citronellic acid and choline: glutaric acid is taken from
FIGS. 6A-6B. NOSEY data for other ILs is shown in SI. Transport
rankings and abbreviations are in Table 2.
[0026] FIG. 8 depicts the range of quaternary ammoniums synthesized
as cationic alternatives to choline.
[0027] FIG. 9A depicts the transport of ruxolitinib (1 mg/ml) to
different portions of the skin over a 24-hour period by ionic
liquid (L-R where horizontal stripes are C1: citronellic acid,
diagonal stripes are C1: geranaic acid, solid black is CAGE, dotted
is choline: citronellic acid). E is epidermis, D is dermis, A is
acceptor fluid. Error bars are SEM (n=3). FIG. 9B depicts 2D NOESY
spectrum of C1: citronellic acid showing one cross-peak
(circled).
[0028] FIG. 10 depicts the transport of ruxolitinib (1 mg/ml) to
different portions of the skin over a 24-hour period by PBS (white
column, none detected) and CAGE ionic liquid (2:1 is shown as
diagonal stripes, 1:1 is solid black, 1:2 is horizontal stripes,
and 1:4 is dotted). E is epidermis, D is dermis, A is acceptor
fluid. Error bars are SE of the mean. Note that pure methanol was
used to extract ruxolitinib from the skin, which also extracted
significant SC lipids simultaneously, dwarfing the drug peak seen
by HPLC. The SC data was thus excluded from the ruxolitinib
analysis.
[0029] FIG. 11 depicts transport of acarbose (1 mg/ml) to different
portions of the skin over a 24-hour period by PBS (white column,
none detected) and CAGE ionic liquid (2:1 is shown as diagonal
stripes, 1:1 is solid black, 1:2 is horizontal stripes, and 1:4 is
dotted). SC is stratum corneum, E is epidermis, D is dermis, A is
acceptor fluid. Error bars are SE of the mean.
[0030] FIG. 12 depicts transport of acarbose to different portions
of the skin over a 24-hour period by ionic liquid (L-R where solid
black is CAGE, dotted is choline citronellate, diagonal downward is
choline hexanoate, wave is choline decanoate, horizontal line is
choline salicylate, weave is choline glutarate, confetti is choline
glycolate, diagonal upward is choline octenoate, and grid is
choline octanoate). SC is stratum corneum, E is epidermis, D is
dermis, A is acceptor fluid. Error bars are SE of the mean.
[0031] FIG. 13 depicts transport of ruxolitinib (1 mg/ml) to
different portions of the skin over a 24-hour period by ionic
liquid (L-R where solid black is CAGE, horizontal line is choline
salicylate, weave is choline glutarate, confetti is choline
glycolate, dotted is choline citronellate, diagonal downward is
choline hexanoate, wave is choline decanoate, diagonal upward is
choline octenoate, and grid is choline octanoate). E is epidermis,
D is dermis, A is acceptor fluid. Error bars are SE of the
mean.
[0032] FIG. 14 depicts 2D NOESY spectrum of Choline Octenoate
(1:2).
[0033] FIG. 15 depicts 2D NOESY spectrum of Choline Octanoate
(1:2).
[0034] FIG. 16 depicts 2D NOESY spectrum of Choline Decanoate
(1:2).
[0035] FIG. 17 depicts 2D NOESY spectrum of Choline Salicylate
(1:2).
[0036] FIG. 18 depicts 2D NOESY spectrum of Choline Glycolate
(1:2).
[0037] FIG. 19 depicts 2D NOESY spectrum of Choline hexanoate
(1:2).
[0038] FIG. 20 depicts 2D NOESY spectrum of Choline geranate (CAGE,
1:2) (from Tanner et al., 2018).
[0039] FIG. 21 depicts 2D NOESY spectrum of Choline alternative 1
with geranic acid (1:2).
[0040] FIG. 22 depicts intestinal administration of insulin with
the indicated ionic liquids. Inulin was mixed with ionic liquids
and administered in the intestine. Blood glucose concentrations and
plasma insulin concentrations were measured. As used in the legend
of FIG. 22, "C--" refers to choline as the cation in an ionic
liquid.
DETAILED DESCRIPTION
[0041] The data provided herein demonstrate that the anion of an
ionic liquid (IL) exerts the predominant influence on whether an
active agent will be transported across a biological barrier (e.g.,
an epithelial layer, such as the dermis). Anions with higher
hydrophobicity and higher mass will provide improved drug delivery
characteristics than anions with lower hydrophobicity and lower
mass. In selecting a cation to pair with the anion, the primary
concern is that the cation not associate too closely with the
anion--close association causes the anion to be retained on the
initial side of the biological barrier.
[0042] Accordingly, in one aspect of any of the embodiments,
described herein is an ionic liquid comprising: i) a hydrophobic
anion comprising a carboxylic acid having a pKa of at least 4.0
and/or a Log P of at least 1.0; and ii) a cation comprising a
quaternary ammonium. In one aspect of any of the embodiments,
described herein is an active compound in combination with at least
one ionic liquid, the ionic liquid comprising: i) a hydrophobic
anion comprising a carboxylic acid having a pKa of at least 4.0
and/or a Log P of at least 1.0; and ii) a cation comprising a
quaternary ammonium. Accordingly, in one aspect of any of the
embodiments, described herein is a method of administering at least
one active compound, the method comprising administering the active
compound in combination with at least one ionic liquid, the ionic
liquid comprising: i) a hydrophobic anion comprising a carboxylic
acid having a pKa of at least 4.0 and/or a Log P of at least 1.0;
and ii) a cation comprising a quaternary ammonium.
[0043] The term "ionic liquids (ILs)" as used herein refers to
organic salts or mixtures of organic salts which are in liquid
state at room temperature. This class of solvents has been shown to
be useful in a variety of fields, including in industrial
processing, catalysis, pharmaceuticals, and electrochemistry. The
ionic liquids contain at least one anionic and at least one
cationic component. Ionic liquids can comprise an additional
hydrogen bond donor (i.e. any molecule that can provide an --OH or
an --NH group), examples include but are not limited to alcohols,
fatty acids, and amines. The at least one anionic and at least one
cationic component may be present in any molar ratio. Exemplary
molar ratios (cation:anion) include but are not limited to 1:1,
1:2, 2:1, 1:3, 3:1, 2:3, 3:2, and ranges between these ratios. For
further discussion of ionic liquids, see, e.g., Hough, et ah, "The
third evolution of ionic liquids: active pharmaceutical
ingredients", New Journal of Chemistry, 31: 1429 (2007) and Xu, et
al., "Ionic Liquids: Ion Mobilities, Glass Temperatures, and
Fragilities", Journal of Physical Chemistry B, 107(25): 6170-6178
(2003); each of which is incorporated by reference herein in its
entirety. In some embodiments of any of the aspects, the ionic
liquid or solvent exists as a liquid below 100.degree. C. In some
embodiments of any of the aspects, the ionic liquid or solvent
exists as a liquid at room temperature.
[0044] As demonstrated herein, anions with higher hydrophobicity
and higher mass will provide improved drug delivery characteristics
than anions with lower hydrophobicity and lower mass. In some
embodiments of any of the aspects, the anion of an IL described
herein is hydrophobic. In some embodiments of any of the aspects,
the anion of an IL described herein is comprises a carboxylic
acid.
[0045] In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 4.0, e.g., 4.0 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 4.5, e.g., 4.5 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 5.0, e.g., 5.0 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at about least 4.0, e.g., about
4.0 or greater. In some embodiments of any of the aspects, the
anion of an IL described herein is has a pKa of at least about 4.5,
e.g., about 4.5 or greater. In some embodiments of any of the
aspects, the anion of an IL described herein is has a pKa of at
least about 5.0, e.g., about 5.0 or greater.
[0046] In some embodiments of any of the aspects, the anion has a
pKa of at least 4.895. In some embodiments of any of the aspects,
the anion has a pKa of 4.5-5.5. In some embodiments of any of the
aspects, the anion has a pKa of 4.895-5.19.
[0047] In some embodiments of any of the aspects, the anion has a
pKa of at least about 4.895. In some embodiments of any of the
aspects, the anion has a pKa of about 4.5 to about 5.5. In some
embodiments of any of the aspects, the anion has a pKa of about
4.895 to about 5.19.
[0048] Hydrophobicity may be assessed by analysis of log P. "Log P"
refers to the logarithm of P (Partition Coefficient). P is a
measure of how well a substance partitions between a lipid (oil)
and water. P itself is a constant. It is defined as the ratio of
concentration of compound in aqueous phase to the concentration of
compound in an immiscible solvent, as the neutral molecule.
[0049] Partition Coefficient, P=[Organic]/[Aqueous] where [
]=concentration
[0050] Log P=log.sub.10 (Partition Coefficient)=log.sub.10 P
In practice, the Log P value will vary according to the conditions
under which it is measured and the choice of partitioning solvent.
A Log P value of 1 means that the concentration of the compound is
ten times greater in the organic phase than in the aqueous phase.
The increase in a log P value of 1 indicates a ten fold increase in
the concentration of the compound in the organic phase as compared
to the aqueous phase.
[0051] In some embodiments of any of the aspects, the anion of an
IL described herein is has a Log P of at least 1.0, e.g., 1.0 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a Log P of at least 2.0, e.g., 2.0 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a Log P of at least 2.5 e.g., 2.5 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a Log P of at least 2.75, e.g., 2.75 or
greater. In some embodiments of any of the aspects, the anion of an
IL described herein is has a Log P of at least about 1.0, e.g.,
about 1.0 or greater. In some embodiments of any of the aspects,
the anion of an IL described herein is has a Log P of at least
about 2.0, e.g., about 2.0 or greater. In some embodiments of any
of the aspects, the anion of an IL described herein is has a Log P
of at least about 2.5 e.g., about 2.5 or greater. In some
embodiments of any of the aspects, the anion of an IL described
herein is has a Log P of at least about 2.75, e.g., about 2.75 or
greater.
[0052] In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 4.0 and a Log P of at
least 1.0. In some embodiments of any of the aspects, the anion of
an IL described herein is has a pKa of at least 4.0 and a Log P of
at least 2.0. In some embodiments of any of the aspects, the anion
of an IL described herein is has a pKa of at least 4.0 and a Log P
of at least 2.5. In some embodiments of any of the aspects, the
anion of an IL described herein is has a pKa of at least 4.0 and a
Log P of at least 2.75.
[0053] In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 4.5 and a Log P of at
least 1.0. In some embodiments of any of the aspects, the anion of
an IL described herein is has a pKa of at least 4.5 and a Log P of
at least 2.0. In some embodiments of any of the aspects, the anion
of an IL described herein is has a pKa of at least 4.5 and a Log P
of at least 2.5. In some embodiments of any of the aspects, the
anion of an IL described herein is has a pKa of at least 4.5 and a
Log P of at least 2.75.
[0054] In some embodiments of any of the aspects, the anion of an
IL described herein is has a pKa of at least 5.0 and a Log P of at
least 1.0. In some embodiments of any of the aspects, the anion of
an IL described herein is has a pKa of at least 5.0 and a Log P of
at least 2.0. In some embodiments of any of the aspects, the anion
of an IL described herein is has a pKa of at least 5.0 and a Log P
of at least 2.5. In some embodiments of any of the aspects, the
anion of an IL described herein is has a pKa of at least 5.0 and a
Log P of at least 2.75.
[0055] In some embodiments of any of the aspects, the anion has a
Log P of at least 2.75. In some embodiments of any of the aspects,
the anion has a Log P of at least 2.8. In some embodiments of any
of the aspects, the anion has a Log P of 2.5-3.5. In some
embodiments of any of the aspects, the anion has a Log P of
2.8-3.01.
[0056] In some embodiments of any of the aspects, the anion has a
Log P of at least about 2.75. In some embodiments of any of the
aspects, the anion has a Log P of at least about 2.8. In some
embodiments of any of the aspects, the anion has a Log P of about
2.5 to about 3.5. In some embodiments of any of the aspects, the
anion has a Log P of about 2.8 to about 3.01.
[0057] In some embodiments of any of the aspects, the carboxylic
acid comprises a carbon backbone chain having 8 carbons and has a
Log P greater than are equal to 2.8 and a pKa between 4.8 and 5.2.
In some embodiments of any of the aspects, the carboxylic acid has
a Log P greater than or equal to 2.9 and a pKa between 4.8 and
5.1.
[0058] The pKa and Log P values for anions are known in the art
and/or can be calculated by one of skill in the art. For example,
PubChem and SpiderChem provide these values for various anions and
chemical manufacturers typically provide them as part of the
catalog listings for their products. pKa and Log P values for
exemplary anions are provided in Table 5 herein.
[0059] In some embodiments of any of the aspects, the carboxylic
acid comprises a carbon chain of at least 6 carbons. In some
embodiments of any of the aspects, the carboxylic acid comprises a
carbon chain of at least 7 carbons. In some embodiments of any of
the aspects, the carboxylic acid comprises a carbon chain of at
least 8 carbons. In some embodiments of any of the aspects, the
carboxylic acid comprises a carbon chain of at least 9 carbons. In
some embodiments of any of the aspects, the carboxylic acid
comprises a carbon chain of at least 10 carbons. In some
embodiments of any of the aspects, the carboxylic acid comprises a
carbon chain of at least 11 carbons.
[0060] In some embodiments of any of the aspects, the anion is an
alkane. In some embodiments of any of the aspects, the anion is an
alkene. In some embodiments of any of the aspects, the anion
comprises a single carboxyl group. In some embodiments of any of
the aspects, the carbon chain of the carboxylic acid comprises one
or more substituent groups. In some embodiments of any of the
aspects, the carbon chain backbone of the carboxylic acid comprises
one or more substituent groups, wherein each substituent group
comprises at least one carbon atom. In some embodiments of any of
the aspects, the carbon chain backbone of the carboxylic acid
comprises one or more substituent groups, wherein at least one
substituent group comprises a methyl group. In some embodiments of
any of the aspects, the carbon chain backbone of the carboxylic
acid comprises two substituent groups, wherein each substituent
group comprises at least one carbon atom. In some embodiments of
any of the aspects, the carbon chain backbone of the carboxylic
acid comprises two substituent groups, wherein one substituent
group comprises a methyl group. In some embodiments of any of the
aspects, the carbon chain backbone of the carboxylic acid comprises
two substituent groups, wherein each substituent group comprises a
methyl group.
[0061] In some embodiments of any of the aspects, the anion is an
unsubstituted alkane. In some embodiments of any of the aspects,
the anion is an unsubstituted alkene. In some embodiments of any of
the aspects, the carbon chain backbone of the carboxylic acid
comprises one or more substituent groups. In some embodiments of
any of the aspects, the carbon chain of the carboxylic acid
comprises one or more substituent groups, wherein each substituent
group comprises at least one carbon atom. In some embodiments of
any of the aspects, the carbon chain of the carboxylic acid
comprises one or more substituent groups, wherein each substituent
group is alkyl, aryl, heteroalkayl, heteroaryl, alkane, or alkene.
In some embodiments of any of the aspects, the carbon chain of the
carboxylic acid comprises one or more substituent groups, wherein
each substituent group is unsubstituted alkyl, unsubstituted aryl,
unsubstituted heteroalkayl, unsubstituted heteroaryl, unsubstituted
alkane, or unsubstituted alkene.
[0062] In some embodiments of any of the aspects, the carboxylic
acid comprises a carbon backbone chain having 8 carbons, is
optionally a mono-alkene, and optionally has two substituents. In
some embodiments of any of the aspects, at least one of the
substituents is a methyl group. In some embodiments of any of the
aspects, both of the substituents is a methyl group. In some
embodiments of any of the aspects, the carboxylic acid is selected
from the group consisting of: octanoic acid; 2-octenoic acid;
3-octenoic acid; 4-octenoic acid; 5-octenoic acid; 6-octenoic acid;
7-octenoic acid; 2,2-dimethyloctanoic acid; 2,3-dimethyloctanoic
acid; 2,4-dimethyloctanoic acid; 2,5-dimethyloctanoic acid;
2,6-dimethyloctanoic acid; 2,7-dimethyloctanoic acid;
3,3-dimethyloctanoic acid; 3,4-dimethyloctanoic acid;
3,5-dimethyloctanoic acid; 3,6-dimethyloctanoic acid;
3,7-dimethyloctanoic acid; 4,4-dimethyloctanoic acid;
4,5-dimethyloctanoic acid; 4,6-dimethyloctanoic acid;
4,7-dimethyloctanoic acid; 5,5-dimethyloctanoic acid;
5,6-dimethyloctanoic acid; 5,7-dimethyloctanoic acid;
6,6-dimethyloctanoic acid; 6,7-dimethyloctanoic acid;
7,7-dimethyloctanoic acid; 2,3-dimethyl-2-octenoic acid;
2,4-dimethyl-2-octenoic acid; 2,5-dimethyl-2-octenoic acid;
2,6-dimethyl-2-octenoic acid; 2,7-dimethyl-2-octenoic acid;
3,4-dimethyl-2-octenoic acid; 3,5-dimethyl-2-octenoic acid;
3,6-dimethyl-2-octenoic acid; 3,7-dimethyl-2-octenoic acid;
4,4-dimethyl-2-octenoic acid; 4,5-dimethyl-2-octenoic acid;
4,6-dimethyl-2-octenoic acid; 4,7-dimethyl-2-octenoic acid;
5,5-dimethyl-2-octenoic acid; 5,6-dimethyl-2-octenoic acid;
5,7-dimethyl-2-octenoic acid; 6,6-dimethyl-2-octenoic acid;
6,7-dimethyl-2-octenoic acid, 7,7-dimethyl-2-octenoic acid;
2,2-dimethyl-3-octenoic acid; 2,3-dimethyl-3-octenoic acid;
2,4-dimethyl-3-octenoic acid; 2,5-dimethyl-3-octenoic acid;
2,6-dimethyl-3-octenoic acid; 2,7-dimethyl-3-octenoic acid;
3,4-dimethyl-3-octenoic acid; 3,5-dimethyl-3-octenoic acid;
3,6-dimethyl-3-octenoic acid; 3,7-dimethyl-3-octenoic acid;
4,5-dimethyl-3-octenoic acid; 4,6-dimethyl-3-octenoic acid;
4,7-dimethyl-3-octenoic acid; 5,5-dimethyl-3-octenoic acid;
5,6-dimethyl-3-octenoic acid; 5,7-dimethyl-3-octenoic acid;
6,6-dimethyl-3-octenoic acid; 6,7-dimethyl-3-octenoic acid;
7,7-dimethyl-3-octenoic acid; 2,2-dimethyl-4-octenoic acid;
2,3-dimethyl-4-octenoic acid; 2,4-dimethyl-4-octenoic acid;
2,5-dimethyl-4-octenoic acid; 2,6-dimethyl-4-octenoic acid;
2,7-dimethyl-4-octenoic acid; 3,3-dimethyl-4-octenoic acid;
3,4-dimethyl-4-octenoic acid; 3,5-dimethyl-4-octenoic acid;
3,6-dimethyl-4-octenoic acid; 3,7-dimethyl-4-octenoic acid;
4,5-dimethyl-4-octenoic acid; 4,6-dimethyl-4-octenoic acid;
4,7-dimethyl-4-octenoic acid; 5,6-dimethyl-4-octenoic acid;
5,7-dimethyl-4-octenoic acid; 6,6-dimethyl-4-octenoic acid;
6,7-dimethyl-4-octenoic acid; 7,7-dimethyl-4-octenoic acid;
2,2-dimethyl-5-octenoic acid; 2,3-dimethyl-5-octenoic acid;
2,4-dimethyl-5-octenoic acid; 2,5-dimethyl-5-octenoic acid;
2,6-dimethyl-5-octenoic acid; 2,7-dimethyl-5-octenoic acid;
3,3-dimethyl-5-octenoic acid; 3,4-dimethyl-5-octenoic acid;
3,5-dimethyl-5-octenoic acid; 3,6-dimethyl-5-octenoic acid;
3,7-dimethyl-5-octenoic acid; 4,4-dimethyl-5-octenoic acid;
4,5-dimethyl-5-octenoic acid; 4,6-dimethyl-5-octenoic acid;
4,7-dimethyl-5-octenoic acid; 5,6-dimethyl-5-octenoic acid;
5,7-dimethyl-5-octenoic acid; 6,7-dimethyl-5-octenoic acid;
7,7-dimethyl-5-octenoic acid; 2,2-dimethyl-6-octenoic acid;
2,3-dimethyl-6-octenoic acid; 2,4-dimethyl-6-octenoic acid;
2,5-dimethyl-6-octenoic acid; 2,6-dimethyl-6-octenoic acid;
2,7-dimethyl-6-octenoic acid; 3,3-dimethyl-6-octenoic acid;
3,4-dimethyl-6-octenoic acid; 3,5-dimethyl-6-octenoic acid;
3,6-dimethyl-6-octenoic acid; 3,7-dimethyl-6-octenoic acid
(citranellic acid); 4,4-dimethyl-6-octenoic acid;
4,5-dimethyl-6-octenoic acid; 4,6-dimethyl-6-octenoic acid;
4,7-dimethyl-6-octenoic acid; 5,5-dimethyl-6-octenoic acid;
5,6-dimethyl-6-octenoic acid; 5,7-dimethyl-6-octenoic acid;
6,7-dimethyl-6-octenoic acid; 2,2-dimethyl-7-octenoic acid;
2,3-dimethyl-7-octenoic acid; 2,4-dimethyl-7-octenoic acid;
2,5-dimethyl-7-octenoic acid; 2,6-dimethyl-7-octenoic acid;
2,7-dimethyl-7-octenoic acid; 4,4-dimethyl-7-octenoic acid;
3,4-dimethyl-7-octenoic acid; 3,5-dimethyl-7-octenoic acid;
3,6-dimethyl-7-octenoic acid; 3,7-dimethyl-7-octenoic acid;
4,4-dimethyl-7-octenoic acid; 4,5-dimethyl-7-octenoic acid;
4,6-dimethyl-7-octenoic acid; 4,7-dimethyl-7-octenoic acid;
5,5-dimethyl-7-octenoic acid; 5,6-dimethyl-7-octenoic acid;
5,7-dimethyl-7-octenoic acid; 6,6-dimethyl-7-octenoic acid;
6,7-dimethyl-7-octenoic acid; and isomers thereof. In some
embodiments of any of the aspects, the carboxylic acid is selected
from the group consisting of: octanoic acid; 2-octenoic acid;
3-octenoic acid; 4-octenoic acid; 5-octenoic acid; 6-octenoic acid;
7-octenoic acid; 2,2-dimethyloctanoic acid; 2,4-dimethyloctanoic
acid; 2,5-dimethyloctanoic acid; 2,6-dimethyloctanoic acid;
2,7-dimethyloctanoic acid; 3,3-dimethyloctanoic acid;
3,5-dimethyloctanoic acid; 3,6-dimethyloctanoic acid;
3,7-dimethyloctanoic acid; 4,4-dimethyloctanoic acid;
4,5-dimethyloctanoic acid; 4,6-dimethyloctanoic acid;
5,5-dimethyloctanoic acid; 5,6-dimethyloctanoic acid;
5,7-dimethyloctanoic acid; 6,6-dimethyloctanoic acid;
7,7-dimethyloctanoic acid; 3,7-dimethyl-2-octenoic acid;
3,7-dimethyl-3-octenoic acid; 3,7-dimethyl-4-octenoic acid;
2,7-dimethyl-6-octenoic acid; 3,7-dimethyl-6-octenoic acid
(citranellic acid); 2,2-dimethyl-7-octenoic acid;
2,3-dimethyl-7-octenoic acid; and isomers thereof. In some
embodiments of any of the aspects, the carboxylic acid is selected
from the group consisting of citranellic acid, octanoic acid,
2-octenoic acid and isomers thereof. In some embodiments of any of
the aspects, the carboxylic acid is selected from the group
consisting of citranellic acid, octanoic acid or trans-2-octenoic
acid. In some embodiments of any of the aspects, octenoic acid as
used herein (for example in Table 5) refers to trans-2-octenoic
acid.
[0063] In some embodiments of any of the aspects, the carboxylic
acid comprises a carbon backbone chain having 8 carbons and is
optionally a mono-alkene. In some embodiments of any of the
aspects, the carbon backbone chain of the carboxylic acid is not
substituted. In some embodiments of any of the aspects, the
carboxylic acid is selected from the group consisting of octanoic
acid, 2-octenoic acid, 3-octenoic acid, 4-octenoic acid, 5-octenoic
acid, 6-octenoic acid, 7-octenoic acid and isomers thereof. In some
options, the carboxylic acid is octanoic acid or trans-2-octenoic
acid (octenoic acid).
[0064] Exemplary, non-limiting anions are provided in Table 5
below.
TABLE-US-00001 TABLE 5 LogP pKa Group 1 Geranic Acid 2.72 5.26
Citronellic Acid 2.8 5.19 Octenoic Acid 2.9 5.05 Decenoic Acid 4.02
5.03 (9Z)-octadec-9-enoic acid 6.5 5.02 Group 2 Octanoic Acid 3.01
4.895 Decanoic Acid 4.09 4.9 (9Z, 12Z)-octadeca-9,12- 7.05 4.77
dienoic acid (R)-5-(1,2-dithiolan-3- 2.1 5.10 yl)pentanoic acid
Group 3 Hexenoic Acid 1.8 5.13 Group 4 Hexanoic Acid 1.92 4.88
3-methylbutanoic acid 1.2 4.77 Nonanedioic Acid 1.57 4.55 Pentanoic
acid 1.39 4.84 Group 5 2-hydroxyoctanoic acid 1.8 4.42
(E)-3-(4-hydroxy-3-methoxy- 1.51 4.42 phenyl)prop-2-enoic acid
Group 6 2-ethylhexyl sulfate 3.10 2-(dimethylamino)ethanol -0.55
9.3 Group 7 8-hydroxycapric acid 2.2 2-methylpropanoic acid 0.73
4.84 Ascorbic Acid -1.85 4.7 Butanoic acid 0.79 4.82 Salicylic Acid
2.2 2.97 Group 8 Hydroxyl(phenyl)acetic acid 1.2 3.41 Glutaric Acid
-0.29 4.34 Adipic acid 0.08 4.4 Group 9 Octanoic Acid 3.01 4.895
Citronellic Acid 2.8 5.19 Octenoic Acid 2.9 5.05 Group 10 Octanoic
Acid 3.01 4.895 Octenoic Acid 2.9 5.05
[0065] In some embodiments of any of the aspects, the anion is
selected from Table 5. In some embodiments of any of the aspects,
the anion is selected from Group 1 of Table 5. In some embodiments
of any of the aspects, the anion is selected from Group 2 of Table
5. In some embodiments of any of the aspects, the anion is selected
from Group 3 of Table 5. In some embodiments of any of the aspects,
the anion is selected from Group 4 of Table 5. In some embodiments
of any of the aspects, the anion is selected from Group 5 of Table
5. In some embodiments of any of the aspects, the anion is selected
from Group 6 of Table 5. In some embodiments of any of the aspects,
the anion is selected from Group 7 of Table 5. In some embodiments
of any of the aspects, the anion is selected from Group 8 of Table
5. In some embodiments of any of the aspects, the anion is selected
from Groups 1-2 of Table 5. In some embodiments of any of the
aspects, the anion is selected from Groups 1-3 of Table 5. In some
embodiments of any of the aspects, the anion is selected from
Groups 1-4 of Table 5. In some embodiments of any of the aspects,
the anion is selected from Groups 1-5 of Table 5. In some
embodiments of any of the aspects, the anion is selected from
Groups 1-6 of Table 5. In some embodiments of any of the aspects,
the anion is selected from Groups 1-7 of Table 5. In some
embodiments of any of the aspects, the anion is selected from Group
9 of Table 5. In some embodiments of any of the aspects, the anion
is selected from Group 10 of Table 5. In some embodiments of any of
the aspects, the anion is selected from Groups 9-10 of Table 5.
[0066] In some embodiments of any of the aspects, the anion is
geranic acid, octanoic acid, and/or citronellic acid. In some
embodiments of any of the aspects, the anion is geranic acid. In
some embodiments of any of the aspects, the anion is octanoic acid.
In some embodiments of any of the aspects, the anion is citronellic
acid. In some embodiments of any of the aspects, the anion
comprises a carbon chain with an 8 carbon backbone. In some
embodiments of any of the aspects, the anion is geranic acid,
octenoic acid, octanoic acid, or citronellic acid. In some
embodiments of any of the aspects, the anion is octenoic acid,
octanoic acid, or citronellic acid.
[0067] As described herein, in selecting a cation to pair with the
anion, the primary concern is that the cation not associate too
closely with the anion--close association causes the anion to be
retained on the initial side of the biological barrier. Choline and
derivatives thereof are shown to be particularly well suited as IL
cations for the types of anions described herein. Accordingly, the
cation of an IL described herein can be a cation comprising a
quaternary ammonium. A quarternary ammonion is a positively charged
polyatomic ion of the structure NR.sub.4.sup.+, each R
independently being an alkyl group or an aryl group.
[0068] In some embodiments of any of the aspects, the cation has a
molar mass equal to or greater than choline, e.g., a molar mass
equal to or greater than 104.1708 g/mol. In some embodiments of any
of the aspects, the cation has a molar mass greater than choline,
e.g., a molar mass equal greater than 104.1708 g/mol.
[0069] In some embodiments of any of the aspects, each R group of
the quarternary ammonium independently comprises an alkyl, alkane,
alkene, or aryl. In some embodiments of any of the aspects, each R
group of the quarternary ammonium independently comprises an alkyl,
alkane, or alkene. In some embodiments of any of the aspects, each
R group of the quarternary ammonium independently comprises an
alkane or alkene. In some embodiments of any of the aspects, each R
group of the quaternernary ammonium independently comprises a
carbon chain of no more than 10 carbon atoms in length, e.g., no
more than 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, or 30
carbon atoms in length. In some embodiments of any of the aspects,
each R group of the quaternernary ammonium independently comprises
a carbon chain of no more than 12 carbon atoms in length. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises a carbon chain of no
more than 15 carbon atoms in length. In some embodiments of any of
the aspects, each R group of the quaternernary ammonium
independently comprises a carbon chain of no more than 20 carbon
atoms in length.
[0070] In some embodiments of any of the aspects, each R group of
the quaternernary ammonium independently comprises a carbon chain
of no more than 10 carbon atoms, e.g., no more than 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, 20, 25, or 30 carbon atoms. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises a carbon chain of no
more than 12 carbon atoms. In some embodiments of any of the
aspects, each R group of the quaternernary ammonium independently
comprises a carbon chain of no more than 15 carbon atoms. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises a carbon chain of no
more than 20 carbon atoms.
[0071] In some embodiments of any of the aspects, each R group of
the quaternernary ammonium independently comprises an alkyl group
of no more than 10 carbon atoms, e.g., no more than 10, 11, 12, 13,
14, 15, 16, 17, 18, 19, 20, 25, or 30 carbon atoms. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises an alkyl group of no
more than 12 carbon atoms. In some embodiments of any of the
aspects, each R group of the quaternernary ammonium independently
comprises an alkyl group of no more than 15 carbon atoms. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises an alkyl group of no
more than 20 carbon atoms.
[0072] In some embodiments of any of the aspects, each R group of
the quaternernary ammonium independently comprises an alkane,
alkene, aryl, heteroaryl, alkyl, or heteroalkyl. In some
embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises an unsubstituted
alkane, unsubstituted alkene, unsubstituted aryl, unsubstituted
heteroaryl, unsubstituted alkyl, or unsubstituted heteroalkyl. In
some embodiments of any of the aspects, each R group of the
quaternernary ammonium independently an unsubstituted alkane. In
some embodiments of any of the aspects, each R group of the
quaternernary ammonium independently an unsubstituted alkene. In
some embodiments of any of the aspects, each R group of the
quaternernary ammonium independently comprises one or more
substituent groups.
[0073] In some embodiments of any of the aspects, at least one R
group of the quaternary ammonium comprises a hydroxy group. In some
embodiments of any of the aspects, one R group of the quaternary
ammonium comprises a hydroxy group. In some embodiments of any of
the aspects, only one R group of the quaternary ammonium comprises
a hydroxy group.
[0074] Exemplary, non-limiting cations can include choline and any
of the cations designated C1-C7 which are defined by structure
below.
##STR00001##
[0075] Further non-limiting examples of cations include the
following: 1-(hydroxymethyl)-1-methylpyrrolidin-1-ium
1-(2-hydroxyethyl)-1-methylpyrrolidin-1-ium
1-ethyl-1-(3-hydroxypropyl)pyrrolidin-1-ium
1-(3-hydroxypropyl)-1-methylpyrrolidin-1-ium
1-(4-hydroxybutyl)-1-methylpyrrolidin-1-ium
1-ethyl-1-(4-hydroxybutyl)pyrrolidin-1-ium
1-(4-hydroxybutyl)-1-propylpyrrolidin-1-ium
1-(5-hydroxypentyl)-1-propylpyrrolidin-1-ium
1-ethyl-1-(5-hydroxypentyl)pyrrolidin-1-ium
1-(5-hydroxypentyl)-1-methylpyrrolidin-1-ium
1-(hydroxymethyl)-1-methylpiperidin-1-ium
1-(2-hydroxyethyl)-1-methylpiperidin-1-ium
1-ethyl-1-(2-hydroxyethyl)piperidin-1-ium
1-ethyl-1-(3-hydroxypropyl)piperidin-1-ium
1-(3-hydroxypropyl)-1-propylpiperidin-1-ium
1-(3-hydroxypropyl)-1-methylpiperidin-1-ium
1-(4-hydroxybutyl)-1-methylpiperidin-1-ium
1-ethyl-1-(4-hydroxybutyl)piperidin-1-ium
1-(4-hydroxybutyl)-1-propylpiperidin-1-ium
I-butyl-1-(5-hydroxypentyl)piperidin-1-ium
1-(5-hydroxyl)entyl)-1-propylpiperidin-1-ium
I-ethyl-1-(5-hydroxypentyl)piperidin-1-ium
1-(5-hydroxyl)entyl)-1-methylpiperidin-1-ium
3-ethyl-1-methyl-1H-imidazol-3-ium
1-methyl-3-propyl-1H-imidazol-3-ium
3-butyl-1-methyl-1H-imidazol-3-ium
1-methyl-3-pentyl-1H-imidazol-3-ium
1,2-dimethyl-3-pentyl-1H-imidazol-3-ium
3-butyl-1,2-dimethyl-1H-imidazol-3-ium
1,2-dimethyl-3-propyl-1H-imidazol-3-ium
3-(hydroxymethyl)-1,2-dimethyl-1H-imidazol-3-ium
3-(2-hydroxyethyl)-1,2-dimethyl-1H-imidazol-3-ium
3-(3-hydroxypropyl)-1,2-dimethyl-1H-imidazol-3-ium
3-(4-hydroxybutyl)-1,2-dimethyl-1H-imidazol-3-ium
3-(5-hydroxypentyl)-1,2-dimethyl-1H-imidazol-3-ium
3-(5-hydroxypentyl)-1-methyl-1H-imidazol-3-ium
3-(4-hydroxybutyl)-1-methyl-1H-imidazol-3-ium
3-(3-hydroxypropyl)-1-methyl-1H-imidazol-3-ium
3-(2-hydroxyethyl)-1-methyl-1H-imidazol-3-ium
3-(hydroxymethyl)-1,2,4,5-tetramethyl-1H-imidazol-3-ium
3-(2-hydroxyethyl)-1,2,4,5-tetramethyl-1H-imidazol-3-ium
3-(3-hydroxypropyl)-1,2,4,5-tetramethyl-1H-imidazol-3-ium
3-(4-hydroxybutyl)-1,2,4,5-tetramethyl-1H-imidazol-3-ium
3-(5-hydroxypentyl)-1,2,4,5-tetramethyl-1H-imidazol-3-ium
1-(5-hydroxypentyl)pyridin-1-ium 1-(4-hydroxybutyl)pyridin-1-ium
1-(3-hydroxypropyl)pyridin-1-ium 1-(2-hydroxyethyl)pyridin-1-ium
1-(hydroxymethyl)pyridin-1-ium 1-hydroxypyridin-1-ium
(hydroxymethyl)trimethylphosphonium
triethyl(hydroxymethyl)phosphonium
triethyl(2-hydroxyethyl)phosphonium
(2-hydroxyethyl)tripropylphosphonium
(3-hydroxypropyl)tripropylphosphonium
tributyl(3-hydroxypropyl)phosphonium
(3-hydroxypropyl)tripentylphosphonium
(4-hydroxybutyl)tripentylphosphonium
(5-hydroxypentyl)tripentylphosphonium
[0076] In some embodiments of any of the aspects, the cation is
choline, C1, C6, and/or C7. In some embodiments of any of the
aspects, the cation is C1, C6, and/or C7.
[0077] In some embodiments of any of the aspects, the cation is
selected from choline, C1, C6, and/or C7 and the anion is
citronellic acid. In some embodiments of any of the aspects, the
cation is choline and the anion is citronellic acid. In some
embodiments of any of the aspects, the cation is C1 and the anion
is citronellic acid. In some embodiments of any of the aspects, the
cation is C6 and the anion is citronellic acid. In some embodiments
of any of the aspects, the cation is C7 and the anion is
citronellic acid.
[0078] In some embodiments of any of the aspects, the cation is
selected from C1, C6, and/or C7 and the anion is geranic acid. In
some embodiments of any of the aspects, the cation is C1 and the
anion is geranic acid. In some embodiments of any of the aspects,
the cation is C6 and the anion is geranic acid. In some embodiments
of any of the aspects, the cation is C7 and the anion is geranic
acid.
[0079] In some embodiments of any of the aspects, the cation is
selected from choline, C1, C6, and/or C7 and the anion is octanoic
acid. In some embodiments of any of the aspects, the cation is
choline and the anion is octanoic acid. In some embodiments of any
of the aspects, the cation is C1 and the anion is octanoic acid. In
some embodiments of any of the aspects, the cation is C6 and the
anion is ocatanoic acid. In some embodiments of any of the aspects,
the cation is C7 and the anion is ocatanoic acid.
[0080] In some embodiments of any of the aspects, the cation is
selected from choline, C1, C6, and C7 and the anion is selected
from citronellic acid, octanoic acid, and octenoic acid. In some
embodiments of any of the aspects, the cation is choline and the
anion is selected from citronellic acid, octanoic acid, and
octenoic acid. In some embodiments of any of the aspects, the ionic
liquid is choline: citronellic acid, choline: octanoic acid, or
choline: octenoic acid.
[0081] In some embodiments of any of the aspects, octenoic acid is
2-octenoic acid.
[0082] Non-limiting, exemplary combinations of cation and anions
are provided in Table 6 below.
TABLE-US-00002 TABLE 6 Choline C1 C2 C3 C4 C5 C6 C7 Group 1 Geranic
Acid X X X X X X X Citronellic Acid X X X X X X X X Octenoic Acid X
X X X X X X X Decenoic Acid X X X X X X X X (9Z)-octadec-9-enoic
acid X X X X X X X X Group 2 Octanoic Acid X X X X X X X X Decanoic
Acid X X X X X X X X (9Z, 12Z)-octadeca-9, X X X X X X X X
12-dienoic acid (R)-5-(1,2-dithiolan-3- X X X X X X X X
yl)pentanoic acid Group 3 Hexenoic Acid X X X X X X X X Group 4
Hexanoic Acid X X X X X X X X 3-methylbutanoic acid X X X X X X X X
Nonanedioic Acid X X X X X X X X Pentanoic acid X X X X X X X X
Group 5 2-hydroxyoctanoic acid X X X X X X X X
(E)-3-(4-hydroxy-3-methoxy- X X X X X X X X phenyl)prop-2-enoic
acid Group 6 2-ethylhexyl sulfate X X X X X X X X
2-(dimethylamino)ethanol X X X X X X X X Group 7 8-hydroxycapric
acid X X X X X X X X 2-methylpropanoic acid X X X X X X X X
Ascorbic Acid X X X X X X X X Butanoic acid X X X X X X X X
Salicylic Acid X X X X X X X X Group 8 Hydroxyl(phenyl)acetic acid
X X X X X X X X Glutaric Acid X X X X X X X X Adipic acid X X X X X
X X X
[0083] In some embodiments of any of the aspects, the ionic liquid
is not CAGE (Choline And GEranate). In some embodiments of any of
the aspects, the cation of the ionic liquid is not choline. In some
embodiments of any of the aspects, the anion of the ionic liquid is
not geranate or geranic acid.
[0084] As demonstrated herein, the number of cross-peaks as
measured by Nuclear Overhauser Effect SpectroscopY (NOESY) for a
given IL correlates with its performance in drug delivery, with
fewer cross peaks indicating better drug delivery performance.
Fundamentally, the number of cross peaks indicates the
intramolecular interactions between the ions that are mediated by
protons. That is, each in-phase peak (the same color as the 1D
diagonal line) indicates that molecules are within 5 nm of each
other as an average across the liquid. In some embodiments of any
of the aspects, the method described herein further comprises a
step of measuring the number of cross-peaks of one or more ILs by
NOESY. In some embodiments of any of the aspects, the ionic liquid
described herein has less than 20 cross peaks as measured by
Nuclear Overhauser Effect SpectroscopY (NOESY), e.g., less than 20,
less than 15, less than 10, less than 9, less than 8, less than 7,
less than 6, or less than 5. In some embodiments of any of the
aspects, the ionic liquid described herein has less than 10 cross
peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY). In some embodiments of any of the aspects, the ionic
liquid described herein has less than 5 cross peaks as measured by
Nuclear Overhauser Effect SpectroscopY (NOESY). Methods of
performing NOESY are known in the art and described in the Examples
herein.
[0085] In one aspect of any of the embodiments, provided herein is
a method of designing, selecting, and/or identifying an ionic
liquid. The ionic liquid can be designed, selected, and/or
identified for administration, administration by a particular route
(e.g., transdermally, to a mucus membrane, orally, subcutaneously,
intradermally, parenterally, intratumorally, or intravenously),
and/or for delivery or administration of one or more active
compounds. As described herein, ionic liquids are most advantageous
for administration, administration by a particular route (e.g.,
transdermally, to a mucus membrane, orally, subcutaneously,
intradermally, parenterally, intratumorally, or intravenously),
and/or for delivery or administration of one or more active
compounds when the inter-ionic interactions of the cation and anion
are reduced. Accordingly, a cation and anion pair can be selected
which minimizes the inter-ionic interactions. For example, the pair
can be selected to minimize the interactions below a threshold
provided herein, to minimize the interactions below a reference
level, and/or to provide the most minimization of any pairwise
combination of cation and anion of cations and anions from a pool
or cations and anions.
[0086] Accordingly, in one aspect of any of the embodiments,
provided herein is a method of designing and/or identifying an
ionic liquid comprising two ions, wherein one ion is a cation and
the other ion is an anion, the method comprising:
in a first option:
[0087] a. selecting one of the two ions of the ionic liquid;
and
[0088] b. selecting the other ion to minimize inter-ionic
interactions;
in a second option:
[0089] a. selecting the cation; and
[0090] b. selecting the anion to minimize inter-ionic interactions;
or
in a third option: [0091] a. selecting the anion; and [0092] b.
selecting the cation to minimize inter-ionic interactions.
[0093] In one aspect of any of the embodiments, provided herein is
a method of designing and/or identifying an ionic liquid comprising
two ions, wherein one ion is a cation and the other ion is an
anion, from a pool of candidate cations and a pool of candidate
anions, the method comprising: in a first option: [0094] a.
selecting one of the two ions of the ionic liquid from the pool of
candidate ions; and [0095] b. selecting from the other pool of
candidate ions the ion which most minimizes inter-ionic
interactions with the ion selected in step a; in a second option:
[0096] a. selecting the cation from the pool of candidate cations;
[0097] b. selecting from the pool of candidate anions the anion
which most minimizes inter-ionic interactions with the cation
selected in step a or in a third option: [0098] a. selecting the
cation from the pool of candidate anions; [0099] b. selecting from
the pool of candidate cations the anion which most minimizes
inter-ionic interactions with the anion selected in step a.
[0100] In some embodiments of any of the aspects, an ionic liquid
with minimized inter-ionic interactions has less than 20 cross
peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY), e.g., less than 20, less than 15, less than 10, less than
9, less than 8, less than 7, less than 6, or less than 5. In some
embodiments of any of the aspects, an ionic liquid with minimized
inter-ionic interactions has less than 10 cross peaks as measured
by Nuclear Overhauser Effect SpectroscopY (NOESY). In some
embodiments of any of the aspects, an ionic liquid with minimized
inter-ionic interactions has less than 5 cross peaks as measured by
Nuclear Overhauser Effect SpectroscopY (NOESY).
[0101] In some embodiments of any of the aspects, the IL is at a
concentration of at least 0.01% w/v. In some embodiments of any of
the aspects, the IL is at a concentration of at least 0.05% w/v. In
some embodiments of any of the aspects, the IL is at a
concentration of at least 0.1% w/v. In some embodiments of any of
the aspects, the IL is at a concentration of at least 0.2% w/v, at
least 0.3% w/v, at least 0.4% w/v, at least 0.5% w/v, at least 1%
w/v or greater. In some embodiments of any of the aspects, the IL
is at a concentration of from about 0.01% w/v to about 1% w/v. In
some embodiments of any of the aspects, the IL is at a
concentration of from 0.01% w/v to 1% w/v. In some embodiments of
any of the aspects, the IL is at a concentration of from about
0.05% w/v to about 0.5% w/v. In some embodiments of any of the
aspects, the IL is at a concentration of from 0.05% w/v to 0.5%
w/v.
[0102] In some embodiments of any of the aspects, the IL is at a
concentration of at least 25% w/w. In some embodiments of any of
the aspects, the IL is at a concentration of at least 25% w/w in
water. In some embodiments of any of the aspects, the IL is at a
concentration of at least 25% w/w in saline or a physiologically
compatible buffer.
[0103] In some embodiments of any of the aspects, the IL is at a
concentration of from about 5% w/w to about 75% w/w. In some
embodiments of any of the aspects, the IL is at a concentration of
from 5% w/w to 75% w/w. In some embodiments of any of the aspects,
the IL is at a concentration of from about 5% w/w to about 75% w/w
in water, saline or a physiologically compatible buffer. In some
embodiments of any of the aspects, the IL is at a concentration of
from 5% w/w to 75% w/w in water, saline or a physiologically
compatible buffer.
[0104] In some embodiments of any of the aspects, the IL is at a
concentration of at least about 0.1% w/w. In some embodiments of
any of the aspects, the IL is at a concentration of at least 0.1%
w/w. In some embodiments of any of the aspects, the IL is at a
concentration of from about 10% w/w to about 70% w/w. In some
embodiments of any of the aspects, the IL is at a concentration of
from 10% w/w to 70% w/w. In some embodiments of any of the aspects,
the IL is at a concentration of from about 30% w/w to about 50%
w/w. In some embodiments of any of the aspects, the IL is at a
concentration of from 30% w/w to 40% w/w. In some embodiments of
any of the aspects, the IL is at a concentration of from about 30%
w/w to about 50% w/w. In some embodiments of any of the aspects,
the IL is at a concentration of from 30% w/w to 40% w/w.
[0105] In some embodiments of any of the aspects, the % w/w
concentration of the IL is % w/w concentration in water, saline, or
a physiologically compatible buffer.
[0106] In some embodiments of any of the aspects, the IL is 100% by
w/w or w/v.
[0107] In some embodiments, the IL is an anhydrous salt, e.g., an
ionic liquid not diluted or dissolved in water. In some
embodiments, the IL is provided as an aqueous solution.
[0108] In some embodiments of any of the aspects, the IL is at a
concentration of at least 25% w/w and has a ratio of cation:anion
of at least 1:3. In some embodiments of any of the aspects, the IL
is at a concentration of at least 25% w/w in water and has a ratio
of cation:anion of at least 1:3. In some embodiments of any of the
aspects, the IL is at a concentration of at least 25% w/w and has a
ratio of cation:anion of 1:3 or 1:4. In some embodiments of any of
the aspects, the IL is at a concentration of at least 25% w/w in
water and has a ratio of cation:anion of 1:3 or 1:4. In some
embodiments of any of the aspects, the IL is a gel, or a
shear-thinning Newtonian gel.
[0109] In some embodiments of any of the aspects, the IL has a
ratio of cation:anion of from about 10:1 to about 1:10. In some
embodiments of any of the aspects, the IL has a ratio of
cation:anion of from 10:1 to 1:10. In some embodiments of any of
the aspects, the IL has a ratio of cation:anion of from about 5:1
to about 1:5. In some embodiments of any of the aspects, the IL has
a ratio of cation:anion of from 5:1 to 1:5. In some embodiments of
any of the aspects, the IL has a ratio of cation:anion of from
about 2:1 to about 1:4. In some embodiments of any of the aspects,
the IL has a ratio of cation:anion of from 2:1 to 1:4. In some
embodiments of any of the aspects, the IL has a ratio of
cation:anion of from about 2:1 to about 1:10. In some embodiments
of any of the aspects, the IL has a ratio of cation:anion of from
2:1 to 1:10. In some embodiments of any of the aspects, the IL has
a ratio of cation:anion such that there is a greater amount of
anion, e.g., a ratio of less than 1:1. In some embodiments of any
of the aspects, the IL has a ratio of cation:anion such that there
is an excess of anion. In some embodiments of any of the aspects,
the IL has a ratio of cation:anion of from about 1:1 to about 1:10.
In some embodiments of any of the aspects, the IL has a ratio of
cation:anion of from 1:1 to 1:10. In some embodiments of any of the
aspects, the IL has a ratio of cation:anion of from about 1:1 to
about 1:4. In some embodiments of any of the aspects, the IL has a
ratio of cation:anion of from 1:1 to 1:4. In some embodiments of
any of the aspects, the IL has a ratio of cation:anion of from
about 1:1 to about 1:3. In some embodiments of any of the aspects,
the IL has a ratio of cation:anion of from 1:1 to 1:3. In some
embodiments of any of the aspects, the IL has a ratio of
cation:anion of from about 1:1 to about 1:2. In some embodiments of
any of the aspects, the IL has a ratio of cation:anion of from 1:1
to 1:2. In some embodiments of any of the aspects, the IL has a
ratio of cation:anion of about 1:1, 1:2, 1:3, or 1:4. In some
embodiments of any of the aspects, the IL has a ratio of
cation:anion of 1:1, 1:2, 1:3, or 1:4. Without wishing to be
constrained by theory, compositions with higher amounts of anion
relative to cation display greater hydrophobicity.
[0110] In some embodiments of any of the aspects, e.g., when one or
more nucleic acid molecules are provided in combination with the
IL, the ratio of cation:anion is greater than 1:1, e.g., greater
than 1:2, from about 1:2 to about 1:4, or from 1:2 to 1:4.
[0111] In some embodiments of any of the aspects, the IL is at a
concentration of at least 20 mM. In some embodiments of any of the
aspects, the IL is at a concentration of at least about 20 mM. In
some embodiments of any of the aspects, the IL is at a
concentration of at least 25 mM. In some embodiments of any of the
aspects, the IL is at a concentration of at least about 25 mM. In
some embodiments of any of the aspects, the IL is at a
concentration of at least 50 mM. In some embodiments of any of the
aspects, the IL is at a concentration of at least about 50 mM. In
some embodiments of any of the aspects, the IL is at a
concentration of at least 100 mM, 500 mM, 1 M, 2 M, 3 M or greater.
In some embodiments of any of the aspects, the IL is at a
concentration of at least about 100 mM, 500 mM, 1 M, 2 M, 3 M or
greater.
[0112] In some embodiments of any of the aspects, the IL is at a
concentration of from about 50 mM to about 4 M. In some embodiments
of any of the aspects, the IL is at a concentration of from 50 mM
to 4 M. In some embodiments of any of the aspects, the IL is at a
concentration of from about 500 mM to about 4 M. In some
embodiments of any of the aspects, the IL is at a concentration of
from 500 mM to 4 M. In some embodiments of any of the aspects, the
IL is at a concentration of from about 1 M to about 4 M. In some
embodiments of any of the aspects, the IL is at a concentration of
from 1 M to 4 M. In some embodiments of any of the aspects, the IL
is at a concentration of from about 2 M to about 4 M. In some
embodiments of any of the aspects, the IL is at a concentration of
from 2 M to 4 M.
[0113] In some embodiments of any of the aspects, the IL
concentration in the composition or formulation is about 0.1 mM to
20 mM. In some embodiments of any of the aspects, the IL
concentration in the composition or formulation is about 0.5 mM to
20 mM, 0.5 mM to 18 mM, 0.5 mM to 16 mM, 0.5 mM to 14 mM, 0.5 mM to
12 mM, 0.5 mM to 10 mM, 0.5 mM to 8 mM, 1 mM to 20 mM, 1 mM to 18
mM, 1 mM to 16 mM, 1 mM to 14 mM, 1 mM to 12 mM, 1 mM to 10 mM, 1
mM to 8 mM, 2 mM to 20 mM, 2 mM to 18 mM, 2 mM to 16 mM, 2 mM to 14
mM, 2 mM to 12 mM, 2 mM to 10 mM, 2 mM to 8 mM, 4 mM to 20 mM, 4 mM
to 18 mM, 4 mM to 16 mM, 4 mM to 14 mM, 4 mM to 12 mM, 4 mM to 10
mM, 4 mM to 8 mM, 6 mM to 20 mM, 6 mM to 18 mM, 6 mM to 14 mM, 6 mM
to 12 mM, 6 mM to 10 mM, 6 mM to 8 mM, 8 mM to 20 mM, 8 mM to 18
mM, 8 mM to 16 mM, 8 mM to 14 mM, 8 mM to 12 mM, 8 mM to 10 mM, 10
mM to 20 mM, 10 mM to 18 mM, 10 mM to 16 mM, 10 mM to 14 mM, 10 mM
to 12 mM, 12 mM to 20 mM, 12 mM to 18 mM, 12 mM to 16 mM, 12 mM to
14 mM, 14 mM to 20 mM, 14 mM to 18 mM, 14 mM to 16 mM, 16 mM to 20
mM, 16 mM to 18 mM, or 18 mM to 20 mM. In some embodiments of any
of the aspects, the IL concentration in the composition or
formulation is about 1 mM, about 2 mM, about 3 mM, about 4 mM,
about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, about
10 mM, about 11 mM, about 12 mM, about 13 mM, about 14 mM, about 15
mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM or about 20
mM.
[0114] It is specifically contemplated that a composition or
combination described herein can comprise one, two, three, or more
of any of the types of components described herein. For example, a
composition can comprise a mixture, solution, combination, or
emulsion of two or more different ionic liquids, and/or a mixture,
solution, combination, or emulsion of two or more different
non-ionic surfactants, and/or a mixture, solution, combination, or
emulsion of two or more different active compounds.
[0115] As used herein, "in combination with" refers to two or more
substances being present in the same formulation in any molecular
or physical arrangement, e.g, in an admixture, in a solution, in a
mixture, in a suspension, in a colloid, in an emulsion. The
formulation can be a homogeneous or heterogenous mixture. In some
embodiments of any of the aspects, the active compound(s) can be
comprised by a superstructure, e.g., nanoparticles, liposomes,
vectors, cells, scaffolds, or the like, said superstructure is
which in solution, mixture, admixture, suspension, etc., with the
IL.
[0116] As used herein, an "active compound" or "active agent" is
any agent which will exert an effect on a target cell or organism.
The terms "compound" and "agent" refer to any entity which is
normally not present or not present at the levels being
administered and/or provided to a cell, tissue or subject. An agent
can be selected from a group comprising: chemicals; small organic
or inorganic molecules; signaling molecules; nucleic acid
sequences; nucleic acid analogues; proteins; peptides; enzymes;
aptamers; peptidomimetic, peptide derivative, peptide analogs,
antibodies; intrabodies; biological macromolecules, extracts made
from biological materials such as bacteria, plants, fungi, or
animal cells or tissues; naturally occurring or synthetic
compositions or functional fragments thereof. In some embodiments,
the agent is any chemical, entity or moiety, including without
limitation synthetic and naturally-occurring non-proteinaceous
entities. Agents can be known to have a desired activity and/or
property, or can be selected from a library of diverse compounds.
Non-limiting examples of active compounds contemplated for use in
the methods described herein include small molecules, polypeptides,
nucleic acids, chemotherapies/chemotherapeutic compounds,
antibodies, antibody reagents, vaccines, a GLP-1 polypeptide or
mimetic/analog thereof, insulin, acarbose, or ruxolitinib.
[0117] A nucleic acid molecule, as described herein, can be a
vector, an expression vector, an inhibitory nucleic acid, an
aptamer, a template molecule or cassette (e.g., for gene editing),
or a targeting molecule (e.g., for CRISPR-Cas technologies), or any
other nucleic acid molecule that one wishes to deliver to a cell.
The nucleic acid molecule can be RNA, DNA, or synthetic or modified
versions thereof.
[0118] In one aspect of any of the embodiments, described herein is
a method of delivering a nucleic acid molecule to a cell, the
method comprising contacting the cell with the nucleic acid
molecule in combination with an IL as described herein. In some
embodiments of any of the aspects, the cell is a cell in a subject
and the contacting step comprises administering the nucleic acid
molecule in combination with the IL to the subject. In some
embodiments of any of the aspects, the cell is in vitro, in vivo,
or ex vivo. In some embodiments of any of the aspects, the cell is
eurkaryotic. In some embodiments of any of the aspects, the cell is
mammalian. In some embodiments of any of the aspects, the cell is
an epithelial cell, e.g, an intestinal epithelial cell.
[0119] As used herein, the term "small molecule" refers to a
chemical agent which can include, but is not limited to, a peptide,
a peptidomimetic, an amino acid, an amino acid analog, a
polynucleotide, a polynucleotide analog, an aptamer, a nucleotide,
a nucleotide analog, an organic or inorganic compound (i.e.,
including heteroorganic and organometallic compounds) having a
molecular weight less than about 10,000 grams per mole, organic or
inorganic compounds having a molecular weight less than about 5,000
grams per mole, organic or inorganic compounds having a molecular
weight less than about 1,000 grams per mole, organic or inorganic
compounds having a molecular weight less than about 500 grams per
mole, and salts, esters, and other pharmaceutically acceptable
forms of such compounds.
[0120] In some embodiments of any of the aspects, the active
compound can be a therapeutic compound or drug, e.g., an agent or
compound which is therapeutically effective for the treatment of at
least one condition in a subject. Therapeutic compounds are known
in the art for a variety of conditions, see, e.g., the database
available on the world wide web at drugs.com or the catalog of
FDA-approved compounds available on the world wide web at
catalog.data.gov/dataset/drugsfda-database; each of which is
incorporated by reference herein in its entirety.
[0121] As used herein the term "chemotherapeutic agent" refers to
any chemical or biological agent with therapeutic usefulness in the
treatment of diseases characterized by abnormal cell growth. Such
diseases include tumors, neoplasms and cancer as well as diseases
characterized by hyperplastic growth. These agents can function to
inhibit a cellular activity upon which the cancer cell depends for
continued proliferation. In some aspect of all the embodiments, a
chemotherapeutic agent is a cell cycle inhibitor or a cell division
inhibitor. Categories of chemotherapeutic agents that are useful in
the methods of the invention include alkylating/alkaloid agents,
antimetabolites, hormones or hormone analogs, and miscellaneous
antineoplastic drugs. Most of these agents are directly or
indirectly toxic to cancer cells. In one embodiment, a
chemotherapeutic agent is a radioactive molecule.
[0122] In some embodiments of any of the aspects, the active
compound is a hydrophobic molecule, e.g., estradiol, testosterone,
corticosterone, paclitaxel, doxorubicin, cisplatin, and/or
camptothecin. In some embodiments of any of the aspects, the active
compound is a hydrophilic molecule.
[0123] In some embodiments of any of the aspects, the active
compound is an antibody or antibody reagent. As used herein, the
term "antibody reagent" refers to a polypeptide that includes at
least one immunoglobulin variable domain or immunoglobulin variable
domain sequence and which specifically binds a given antigen. An
antibody reagent can comprise an antibody or a polypeptide
comprising an antigen-binding domain of an antibody. In some
embodiments, an antibody reagent can comprise a monoclonal antibody
or a polypeptide comprising an antigen-binding domain of a
monoclonal antibody. For example, an antibody can include a heavy
(H) chain variable region (abbreviated herein as VH), and a light
(L) chain variable region (abbreviated herein as VL). In another
example, an antibody includes two heavy (H) chain variable regions
and two light (L) chain variable regions. The term "antibody
reagent" encompasses antigen-binding fragments of antibodies (e.g.,
single chain antibodies, Fab and sFab fragments, F(ab')2, Fd
fragments, Fv fragments, scFv, and domain antibodies (dAb)
fragments as well as complete antibodies.
[0124] In some embodiments of any of the aspects, the active
compound has a molecular weight of greater than about 450. In some
embodiments of any of the aspects, the active compound has a
molecular weight of greater than about 500. In some embodiments of
any of the aspects, the active compound has a molecular weight of
greater than 450, e.g., greater than 450, greater than 500, greater
than 550, greater than 600, greater than 1000 or more. In some
embodiments of any of the aspects, the active compound is
polar.
[0125] In some embodiments of any of the aspects, a composition or
combination as described herein, comprising at least one IL and
optionally an active compound can be formulated as an oral,
subcutaneous, intravenous, intradermal, or parenteral formulation.
In some embodiments of any of the aspects, an oral formulation can
be a degradable capsule comprising the composition comprising the
at least one IL and optionally, an active compound.
[0126] In some embodiments of any of the aspects, described herein
is a composition comprising at least one IL as described herein and
at least one active compound. In some embodiments of any of the
aspects, described herein is a composition consisting essentially
of at least one IL as described herein and at least one active
compound. In some embodiments of any of the aspects, described
herein is a composition consisting of at least one IL as described
herein and at least one active compound. In some embodiments of any
of the aspects, the composition comprising at least one IL as
described herein and at least one active compound is administered
as a monotherapy, e.g., another treatment for the condition is not
administered to the subject.
[0127] In one aspect of any of the embodiments, described herein is
a pharmaceutical composition comprising at least one active
compound in combination with at least one IL as described herein.
In some embodiments, the pharmaceutical composition comprises the
at least one IL and the one or more active compounds as described
herein. In some embodiments, the pharmaceutical composition
consists essentially of the at least one IL and the one or more
active compounds as described herein. In some embodiments, the
pharmaceutical composition consists of the at least one IL and the
one or more active compounds as described herein. In some
embodiments, the pharmaceutical composition consists essentially of
an aqeuous solution of the at least one IL and the one or more
active compounds as described herein. In some embodiments, the
pharmaceutical composition consists of an aqeuous solution of the
at least one IL and the one or more active compounds as described
herein.
[0128] The compositions, formulations, and combinations described
herein can comprise at least one IL as described herein, e.g., one
IL, two ILs, three ILs, or more. In some embodiments of any of the
aspects, a composition, formulation, or combination as described
herein can comprise at least one IL as described herein and CAGE
(Choline And GEranate).
[0129] In some embodiments of any of the aspects, the at least one
active compound and the at least one ionic liquid are further in
combination with at least one non-ionic surfactant. As used herein,
"non-ionic surfactant" refers to a surfactant which lacks a net
ionic charge and does not dissociate to an appreciable extent in
aqueous media. The properties of non-ionic surfactants are largely
dependent upon the proportions of the hydrophilic and hydrophobic
groups in the molecule. Hydrophilic groups include the oxyethylene
group (--OCH2 CH2-) and the hydroxy group. By varying the number of
these groups in a hydrophobic molecule, such as a fatty acid,
substances are obtained which range from strongly hydrophobic and
water insoluble compounds, such as glyceryl monostearate, to
strongly hydrophilic and water-soluble compounds, such as the
macrogols. Between these two extremes types include those in which
the proportions of the hydrophilic and hydrophobic groups are more
evenly balanced, such as the macrogol esters and ethers and
sorbitan derivatives. Suitable non-ionic surfactants may be found
in Martindale, The Extra Pharmacopoeia, 28th Edition, 1982, The
Pharmaceutical Press, London, Great Britain, pp. 370 to 379.
Non-limiting examples of non-ionic surfactants include
polysorbates, a Tween.TM., block copolymers of ethylene oxide and
propylene oxide, glycol and glyceryl esters of fatty acids and
their derivatives, polyoxyethylene esters of fatty acids (macrogol
esters), polyoxyethylene ethers of fatty acids and their
derivatives (macrogol ethers), polyvinyl alcohols, and sorbitan
esters, sorbitan monoesters, ethers formed from fatty alcohols and
polyethylene glycol, polyoxyethylene-polypropylene glycol, alkyl
polyglycoside, Cetomacrogol 1000, cetostearyl alcohol, cetyl
alcohol, cocamide DEA, cocamide MEA, decyl glucoside, decyl
polyglucose, glycerol monostearate, IGEPAL CA-630, isoceteth-20,
lauryl glucoside, maltosides, monolaurin, mycosubtilin, Nonidet
P-40, nonoxynol-9, nonoxynols, NP-40, octaethylene glycol
monododecyl ether, N-Octyl beta-D-thioglucopyranoside, octyl
glucoside, oleyl alcohol, PEG-10 sunflower glycerides,
pentaethylene glycol monododecyl ether, polidocanol, poloxamer,
poloxamer 407, polyethoxylated tallow amine, polyglycerol
polyricinoleate, sorbitan, sorbitan monolaurate, sorbitan
monostearate, sorbitan tristearate, stearyl alcohol, surfactin,
Triton X-100, and the like. In some embodiments of any of the
aspects, the at least one non-ionic surfactant has a neutral
hydrophilic head group.
[0130] As used herein, "polysorbate" refers to a surfactant derived
from ethoxylated sorbitan (a derivative of sorbitol) esterified
with fatty acids. Common brand names for polysorbates include
Scattics.TM., Alkest.TM., Canarcel.TM., and Tween.TM.. Exemplary
polysorbates include polysorbate 20 (polyoxyethylene (20) sorbitan
monolaurate), polysorbate 40 (polyoxyethylene (20) sorbitan
monopalmitate), polysorbate 60 (polyoxyethylene (20) sorbitan
monostearate), and polysorbate 80 (polyoxyethylene (20) sorbitan
monooleate).
[0131] In some embodiments of any of the aspects, the at least one
non-ionic surfactant (e.g., at least one polysorbate) is present at
a concentration of about 0.1% to about 50% w/v. In some embodiments
of any of the aspects, the at least one non-ionic surfactant (e.g.,
at least one polysorbate) is present at a concentration of 0.1% to
50% w/v. In some embodiments of any of the aspects, the at least
one non-ionic surfactant (e.g., at least one polysorbate) is
present at a concentration of about 1% to about 5% w/v. In some
embodiments of any of the aspects, the at least one non-ionic
surfactant (e.g., at least one polysorbate) is present at a
concentration of 1% to 5% w/v. In some embodiments of any of the
aspects, the at least one non-ionic surfactant (e.g., at least one
polysorbate) is present at a concentration of about 3% to about 10%
w/v. In some embodiments of any of the aspects, the at least one
non-ionic surfactant (e.g., at least one polysorbate) is present at
a concentration of 3% to 10% w/v. In some embodiments of any of the
aspects, the at least one non-ionic surfactant (e.g., at least one
polysorbate) is present at a concentration of less than about 5%
w/v. In some embodiments of any of the aspects, the at least one
non-ionic surfactant (e.g., at least one polysorbate) is present at
a concentration of less than 5% w/v.
[0132] In some embodiments of any of the aspects, the combination
of the at least one active compound and at least one IL as
described herein is provided in one or more nanoparticles. In some
embodiments of any of the aspects, the combination of the at least
one active compound and at least one IL as described herein
comprises nanoparticles comprising the active compound, the
nanoparticles in solution or suspension in a composition comprising
at least one IL as described herein.
[0133] In some embodiments of any of the aspects, a composition as
described herein, e.g., a composition comprising at least one IL
and an active compound, can further comprise a pharmaceutically
acceptable carrier. As used herein, the terms "pharmaceutically
acceptable", "physiologically tolerable" and grammatical variations
thereof, as they refer to compositions, carriers, diluents and
reagents, are used interchangeably and represent that the materials
are capable of administration to or upon a mammal without the
production of undesirable physiological effects such as nausea,
dizziness, gastric upset and the like. A pharmaceutically
acceptable carrier will not promote the raising of an immune
response to an agent with which it is admixed, unless so desired.
The preparation of a pharmacological composition that contains
active ingredients dissolved or dispersed therein is well
understood in the art and need not be limited based on formulation.
Typically, such compositions are prepared as injectable either as
liquid solutions or suspensions, however, solid forms suitable for
solution, or suspensions, in liquid prior to use can also be
prepared. The preparation can also be emulsified or presented as a
liposome composition. The active ingredient can be mixed with
excipients which are pharmaceutically acceptable and compatible
with the active ingredient and in amounts suitable for use in the
therapeutic methods described herein. Suitable excipients include,
for example, water, saline, dextrose, glycerol, ethanol or the like
and combinations thereof. In addition, if desired, the composition
can contain minor amounts of auxiliary substances such as wetting
or emulsifying agents, pH buffering agents and the like which
enhance the effectiveness of the active ingredient. The therapeutic
composition of the present disclosure can include pharmaceutically
acceptable salts of the components therein. Pharmaceutically
acceptable salts include the acid addition salts (formed with the
free amino groups of the polypeptide) that are formed with
inorganic acids such as, for example, hydrochloric or phosphoric
acids, or such organic acids as acetic, tartaric, mandelic and the
like. Salts formed with the free carboxyl groups can also be
derived from inorganic bases such as, for example, sodium,
potassium, ammonium, calcium or ferric hydroxides, and such organic
bases as isopropylamine, trimethylamine, 2-ethylamino ethanol,
histidine, procaine and the like. Physiologically tolerable
carriers are well known in the art. Exemplary liquid carriers are
sterile aqueous solutions that contain no materials in addition to
the active ingredients and water, or contain a buffer such as
sodium phosphate at physiological pH value, physiological saline or
both, such as phosphate-buffered saline. Still further, aqueous
carriers can contain more than one buffer salt, as well as salts
such as sodium and potassium chlorides, dextrose, polyethylene
glycol and other solutes. Liquid compositions can also contain
liquid phases in addition to and to the exclusion of water.
Exemplary of such additional liquid phases are glycerin, vegetable
oils such as cottonseed oil, and water-oil emulsions. The amount of
an active agent used in the methods described herein that will be
effective in the treatment of a particular disorder or condition
will depend on the nature of the disorder or condition, and can be
determined by standard clinical techniques. Suitable pharmaceutical
carriers are described in Remington's Pharmaceutical Sciences, A.
Osol, a standard reference text in this field of art. For example,
a parenteral composition suitable for administration by injection
is prepared by dissolving 1.5% by weight of active ingredient in
0.9% sodium chloride solution.
[0134] The term "carrier" in the context of a pharmaceutical
carrier refers to a diluent, adjuvant, excipient, or vehicle with
which the therapeutic is administered. Such pharmaceutical carriers
can be sterile liquids, such as water and oils, including those of
petroleum, animal, vegetable or synthetic origin, such as peanut
oil, soybean oil, mineral oil, sesame oil and the like. Water is a
preferred carrier when the pharmaceutical composition is
administered intravenously. Saline solutions and aqueous dextrose
and glycerol solutions can also be employed as liquid carriers,
particularly for injectable solutions. Suitable pharmaceutical
excipients include starch, glucose, lactose, sucrose, gelatin,
malt, rice, flour, chalk, silica gel, sodium stearate, glycerol
monostearate, talc, sodium chloride, dried skim milk, glycerol,
propylene, glycol, water, ethanol and the like. The composition, if
desired, can also contain minor amounts of wetting or emulsifying
agents, or pH buffering agents. These compositions can take the
form of solutions, suspensions, emulsion, tablets, pills, capsules,
powders, sustained-release formulations, and the like. The
composition can be formulated as a suppository, with traditional
binders and carriers such as triglycerides. Oral formulation can
include standard carriers such as pharmaceutical grades of
mannitol, lactose, starch, magnesium stearate, sodium saccharine,
cellulose, magnesium carbonate, etc. Examples of suitable
pharmaceutical carriers are described in Remington's Pharmaceutical
Sciences, 18th Ed., Gennaro, ed. (Mack Publishing Co., 1990). The
formulation should suit the mode of administration.
[0135] Pharmaceutically acceptable carriers and diluents include
saline, aqueous buffer solutions, solvents and/or dispersion media.
The use of such carriers and diluents is well known in the art.
Some non-limiting examples of materials which can serve as
pharmaceutically-acceptable carriers include: (1) sugars, such as
lactose, glucose and sucrose; (2) starches, such as corn starch and
potato starch; (3) cellulose, and its derivatives, such as sodium
carboxymethyl cellulose, methylcellulose, ethyl cellulose,
microcrystalline cellulose and cellulose acetate; (4) powdered
tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as
magnesium stearate, sodium lauryl sulfate and talc; (8) excipients,
such as cocoa butter and suppository waxes; (9) oils, such as
peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil,
corn oil and soybean oil; (10) glycols, such as propylene glycol;
(11) polyols, such as glycerin, sorbitol, mannitol and polyethylene
glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate;
(13) agar; (14) buffering agents, such as magnesium hydroxide and
aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water;
(17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol;
(20) pH buffered solutions; (21) polyesters, polycarbonates and/or
polyanhydrides; (22) bulking agents, such as polypeptides and amino
acids (23) serum component, such as serum albumin, HDL and LDL;
(22) C.sub.2-C.sub.12 alcohols, such as ethanol; and (23) other
non-toxic compatible substances employed in pharmaceutical
formulations. Wetting agents, coloring agents, release agents,
coating agents, sweetening agents, flavoring agents, perfuming
agents, preservative and antioxidants can also be present in the
formulation. The terms such as "excipient", "carrier",
"pharmaceutically acceptable carrier" or the like are used
interchangeably herein. In some embodiments, the carrier inhibits
the degradation of the active compound. The term "pharmaceutically
acceptable carrier" excludes tissue culture medium.
[0136] In some embodiments of any of the aspects, a composition as
described herein, e.g, a composition comprising at least one IL as
described herein and an active compound, can be formulated as an
oral, subcutaneous, intravenous, intradermal, or parenteral
formulation. In some embodiments of any of the aspects, an oral
formulation can be a degradable capsule comprising the composition
described herein, e.g., a composition comprising at least one IL as
described herein and an active compound.
[0137] In some embodiments of any of the aspects described herein,
the biological activity of the active compound is improved or
stabilized as compared to the activity in the absence of the at
least one IL. In some embodiments of any of the aspects described
herein, the greatly enhances permeation of the active compound
across the skin compared to a control where the at least one IL is
absent.
[0138] In one aspect of any of the embodiments, described herein is
a method of administering at least active compound to a subject
using a catheter wherein the catheter is coated with at least one
IL as described herein. In one aspect of any of the embodiments,
described herein is a method of collecting a body fluid by placing
the catheter into the body wherein the catheter is coated with at
least one IL as described herein.
[0139] In one aspect of any of the embodiments, the composition or
combination described herein is for a method of administering or
delivering at least one active compound, e.g., for the treatment of
a disease. In one aspect of any of the embodiments, described
herein is a method of administering at least one active compound,
the method comprising administering the active compound in
combination with at least one IL as described herein. In one aspect
of any of the embodiments, described herein is a method of treating
a disease by administering at least one active compound, the method
comprising administering the active compound in combination with at
least one IL as described herein.
[0140] In some embodiments, the methods described herein relate to
treating a subject having or diagnosed as having a condition with a
composition as described herein, e.g, a comprising at least one IL
and an active compound. Subjects having a condition, e.g.,
diabetes, can be identified by a physician using current methods of
diagnosing diabetes. Symptoms and/or complications of diabetes
which characterize these conditions and aid in diagnosis are well
known in the art and include but are not limited to, weight loss,
slow healing, polyuria, polydipsia, polyphagiam headaches, itchy
skin, and fatigue. Tests that may aid in a diagnosis of, e.g.
diabetes include, but are not limited to, blood tests (e.g., for
fasting glucose levels). A family history of diabetes, or exposure
to risk factors for diabetes (e.g. overweight) can also aid in
determining if a subject is likely to have diabetes or in making a
diagnosis of diabetes.
[0141] The compositions and methods described herein can be
administered to a subject having or diagnosed as having a condition
described herein. In some embodiments, the methods described herein
comprise administering an effective amount of compositions
described herein, e.g. a composition comprising at least one IL as
described herein and an active compound, to a subject in order to
alleviate a symptom of a condition described herein. As used
herein, "alleviating a symptom" is ameliorating any marker or
symptom associated with a condition. As compared with an equivalent
untreated control, such reduction is by at least 5%, 10%, 20%, 40%,
50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard
technique. A variety of means for administering the compositions
described herein to subjects are known to those of skill in the
art. Such methods can include, but are not limited to oral,
parenteral, intravenous, intramuscular, subcutaneous, transdermal,
airway (aerosol), pulmonary, cutaneous, injection, or intratumoral
administration. Administration can be local or systemic.
[0142] In some embodiments of any of the aspects, the
administration is transdermal. In some embodiments of any of the
aspects, the administration is transdermal, to a mucus membrane
(e.g., to a nasal, oral, or vaginal membrane), oral, subcutaneous,
intradermal, parenteral, intratumoral, or intravenous.
[0143] Oral administration can comprise providing tablets
(including without limitation scored or coated tablets), pills,
caplets, capsules, chewable tablets, powder packets, cachets,
troches, wafers, aerosol sprays, or liquids, such as but not
limited to, syrups, elixirs, solutions or suspensions in an aqueous
liquid, a non-aqueous liquid, an oil-in-water emulsion, or a
water-in-oil emulsion. Oral formulations can comprise discrete
dosage forms, such as, but not limited to, tablets (including
without limitation scored or coated tablets), pills, caplets,
capsules, chewable tablets, powder packets, cachets, troches,
wafers, aerosol sprays, or liquids, such as but not limited to,
syrups, elixirs, solutions or suspensions in an aqueous liquid, a
non-aqueous liquid, an oil-in-water emulsion, or a water-in-oil
emulsion. Such compositions contain a predetermined amount of an
ionic liquid as described herein and the at least one active
compound, and may be prepared by methods of pharmacy well known to
those skilled in the art. See generally, Remington: The Science and
Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins,
Philadelphia Pa. (2005).
[0144] In one aspect of any of the embodiments, described herein is
a method of delivery of at least one active compound by
subcutaneous, intradermal or intravenous administration, the method
comprising administering the active compound in combination with at
least one IL as described herein. In some embodiments of any of the
aspects, subcutaneous, intradermal or intravenous administration
comprises administration via injection, catheter, port, or the
like.
[0145] In one aspect of any of the embodiments, described herein is
a method of parenteral delivery of at least one active compound,
the method comprising parenterally administering the active
compound in combination with at least one IL as described herein.
In some embodiments, the parenteral administration comprises
delivery to a tumor, e.g., a cancer tumor. In some embodiments of
any of the aspects, the composition or combination described herein
can be a parenteral dose form. Since administration of parenteral
dosage forms typically bypasses the patient's natural defenses
against contaminants, parenteral dosage forms are preferably
sterile or capable of being sterilized prior to administration to a
patient. Examples of parenteral dosage forms include, but are not
limited to, solutions ready for injection, dry products ready to be
dissolved or suspended in a pharmaceutically acceptable vehicle for
injection, suspensions ready for injection, and emulsions. In
addition, controlled-release parenteral dosage forms can be
prepared for administration of a patient, including, but not
limited to, DUROS.RTM.-type dosage forms and dose-dumping.
[0146] Suitable vehicles that can be used to provide parenteral
dosage forms of a composition comprising an ionic liquid as
described herein in combination with at least one active compound
as disclosed within are well known to those skilled in the art.
Examples include, without limitation: sterile water; water for
injection USP; saline solution; glucose solution; aqueous vehicles
such as but not limited to, sodium chloride injection, Ringer's
injection, dextrose Injection, dextrose and sodium chloride
injection, and lactated Ringer's injection; water-miscible vehicles
such as, but not limited to, ethyl alcohol, polyethylene glycol,
and propylene glycol; and non-aqueous vehicles such as, but not
limited to, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl
oleate, isopropyl myristate, and benzyl benzoate. Compounds that
alter or modify the solubility of an ingredient in a composition as
disclosed herein can also be incorporated into the parenteral
dosage forms of the disclosure, including conventional and
controlled-release parenteral dosage forms.
[0147] Conventional dosage forms generally provide rapid or
immediate drug release from the formulation. Depending on the
pharmacology and pharmacokinetics of the drug, use of conventional
dosage forms can lead to wide fluctuations in the concentrations of
the drug in a patient's blood and other tissues. These fluctuations
can impact a number of parameters, such as dose frequency, onset of
action, duration of efficacy, maintenance of therapeutic blood
levels, toxicity, side effects, and the like. While as noted above
herein, the compositions comprising an ionic liquid as described
herein in combination with at least one active compound can obviate
certain reasons for using a controlled-release formulation, it is
contemplated herein that the methods and compositions can be
utilized in controlled-release formulations in some embodiments.
For example, controlled-release formulations can be used to control
a drug's onset of action, duration of action, plasma levels within
the therapeutic window, and peak blood levels. In particular,
controlled- or extended-release dosage forms or formulations can be
used to ensure that the maximum effectiveness of a drug is achieved
while minimizing potential adverse effects and safety concerns,
which can occur both from under-dosing a drug (i.e., going below
the minimum therapeutic levels) as well as exceeding the toxicity
level for the drug. In some embodiments, the composition comprising
an ionic liquid as described herein in combination with at least
one active compound can be administered in a sustained release
formulation.
[0148] Controlled-release pharmaceutical products have a common
goal of improving drug therapy over that achieved by their
non-controlled release counterparts. Ideally, the use of an
optimally designed controlled-release preparation in medical
treatment is characterized by a minimum of drug substance being
employed to cure or control the condition in a minimum amount of
time. Advantages of controlled-release formulations include: 1)
extended activity of the drug; 2) reduced dosage frequency; 3)
increased patient compliance; 4) usage of less total drug; 5)
reduction in local or systemic side effects; 6) minimization of
drug accumulation; 7) reduction in blood level fluctuations; 8)
improvement in efficacy of treatment; 9) reduction of potentiation
or loss of drug activity; and 10) improvement in speed of control
of diseases or conditions. Kim, Cherng-ju, Controlled Release
Dosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.:
2000).
[0149] Most controlled-release formulations are designed to
initially release an amount of drug (active ingredient) that
promptly produces the desired therapeutic effect, and gradually and
continually release other amounts of drug to maintain this level of
therapeutic or prophylactic effect over an extended period of time.
In order to maintain this constant level of drug in the body, the
drug must be released from the dosage form at a rate that will
replace the amount of drug being metabolized and excreted from the
body. Controlled-release of an active ingredient can be stimulated
by various conditions including, but not limited to, pH, ionic
strength, osmotic pressure, temperature, enzymes, water, and other
physiological conditions or compounds.
[0150] A variety of known controlled- or extended-release dosage
forms, formulations, and devices can be adapted for use with the
salts and compositions of the disclosure. Examples include, but are
not limited to, those described in U.S. Pat. Nos. 3,845,770;
3,916,899; 3,536,809; 3,598,123; 4,008,719; 5,674,533; 5,059,595;
5,591,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,733,566;
and 6,365,185 B1; each of which is incorporated herein by
reference. These dosage forms can be used to provide slow or
controlled-release of one or more active ingredients using, for
example, hydroxypropylmethyl cellulose, other polymer matrices,
gels, permeable membranes, osmotic systems (such as OROS.RTM. (Alza
Corporation, Mountain View, Calif. USA)), or a combination thereof
to provide the desired release profile in varying proportions.
[0151] The term "effective amount" as used herein refers to the
amount of a composition needed to alleviate at least one or more
symptom of the disease or disorder, and relates to a sufficient
amount of pharmacological composition to provide the desired
effect. The term "therapeutically effective amount" therefore
refers to an amount of a composition that is sufficient to provide
a particular effect when administered to a typical subject. An
effective amount as used herein, in various contexts, would also
include an amount sufficient to delay the development of a symptom
of the disease, alter the course of a symptom disease (for example
but not limited to, slowing the progression of a symptom of the
disease), or reverse a symptom of the disease. Thus, it is not
generally practicable to specify an exact "effective amount".
However, for any given case, an appropriate "effective amount" can
be determined by one of ordinary skill in the art using only
routine experimentation.
[0152] Effective amounts, toxicity, and therapeutic efficacy can be
determined by standard pharmaceutical procedures in cell cultures
or experimental animals, e.g., for determining the LD50 (the dose
lethal to 50% of the population) and the ED50 (the dose
therapeutically effective in 50% of the population). The dosage can
vary depending upon the dosage form employed and the route of
administration utilized. The dose ratio between toxic and
therapeutic effects is the therapeutic index and can be expressed
as the ratio LD50/ED50. Compositions and methods that exhibit large
therapeutic indices are preferred. A therapeutically effective dose
can be estimated initially from cell culture assays. Also, a dose
can be formulated in animal models to achieve a circulating plasma
concentration range that includes the IC50 (i.e., the concentration
of the active compound, which achieves a half-maximal inhibition of
symptoms) as determined in cell culture, or in an appropriate
animal model. Levels in plasma can be measured, for example, by
high performance liquid chromatography. The effects of any
particular dosage can be monitored by a suitable bioassay, e.g.,
assay for blood glucose, among others. The dosage can be determined
by a physician and adjusted, as necessary, to suit observed effects
of the treatment.
[0153] As used herein, "diabetes" refers to diabetes mellitus, a
metabolic disease characterized by a deficiency or absence of
insulin secretion by the pancreas. As used throughout, "diabetes"
includes Type 1, Type 2, Type 3, and Type 4 diabetes mellitus
unless otherwise specified herein. The onset of diabetes is
typically due to a combination of hereditary and environmental
causes, resulting in abnormally high blood sugar levels
(hyperglycemia). The two most common forms of diabetes are due to
either a diminished production of insulin (in type 1), or
diminished response by the body to insulin (in type 2 and
gestational). Both lead to hyperglycemia, which largely causes the
acute signs of diabetes: excessive urine production, resulting
compensatory thirst and increased fluid intake, blurred vision,
unexplained weight loss, lethargy, and changes in energy
metabolism. Diabetes can cause many complications. Acute
complications (hypoglycemia, ketoacidosis, or nonketotic
hyperosmolar coma) may occur if the disease is not adequately
controlled. Serious long-term complications (i.e. chronic side
effects) include cardiovascular disease (doubled risk), chronic
renal failure, retinal damage (which can lead to blindness), nerve
damage (of several kinds), and microvascular damage, which may
cause impotence and poor wound healing. Poor healing of wounds,
particularly of the feet, can lead to gangrene, and possibly to
amputation. In some embodiments, the diabetes can be Type 2
diabetes. Type 2 diabetes (non-insulin-dependent diabetes mellitus
(NIDDM), or adult-onset diabetes) is a metabolic disorder that is
primarily characterized by insulin resistance (diminished response
by the body to insulin), relative insulin deficiency, and
hyperglycemia. In some embodiments, a subject can be pre-diabetic,
which can be characterized, for example, as having elevated fasting
blood sugar or elevated post-prandial blood sugar.
[0154] Glucagon-Like Peptide-1(GLP-1), is known to reduce food
intake and hunger feelings in humans and is an incretin derived
from the transcription product of the proglucagon gene that
contributes to glucose homeostasis. GLP-1 mimetics are currently
being used in the treatment of Type 2 diabetes. Recent clinical
trials have shown that these treatments not only improve glucose
homeostasis but also succeed in inducing weight loss. As used
herein. "GLP-1 polypeptide" refers to the various pre- and
pro-peptides and cleavage products of GLP-1, e.g., for human:
GLP-1(1-37) (SEQ ID NO: 2), GLP-1 (7-36) (SEQ ID NO: 3), and GLP-1
(7-37) (SEQ ID NO: 4). In some embodiments, a GLP-1 polypeptide can
be GLP-1 (7-36) and/or GLP-1 (7-37) or the correlating polypeptides
from a species other than human. Sequences for GLP-1 polypeptides
are known in the art for a number of species, e.g. human GLP-1
(NCBI Gene ID: 2641) polypeptides (e.g., NCBI Ref Seq: NP_002045.1;
SEQ ID NO: 1) and SEQ ID NOs: 2-4. In some embodiments, a pre or
pro-peptide of GLP-1 can be used in the methods or compositions
described herein, e.g., a glucagon preproprotein (e.g., SEQ ID NO:
1). Naturally-occurring alleles or variants of any of the
polypeptides described herein are also specifically contemplated
for use in the methods and compositions described herein.
TABLE-US-00003 SEQ ID NO: 1 1 mksiyfvagl fvmlvqgswq rslqdteeks
rsfsasqadp lsdpdqmned krhsqgtfts 61 dyskyldsrr aqdfvqwlmn
tkrnrnniak rhdeferhae gtftsdvssy legqaakefi 121 awlvkgrgrr
dfpeevaive elgrrhadgs fsdemntild nlaardfinw liqtkitdrk SEQ ID NO: 2
hdeferhae gtftsdvssy legqaakefi awlvkgrg SEQ ID NO: 3 hae
gtftsdvssy legqaakefi awlvkgr SEQ ID NO: 4 hae gtftsdvssy
legqaakefi awlvkgrg
[0155] Various GLP-1 mimetics are known in the art and used in the
treatment of diabetes. GLP-1 mimetics (or analogues) can include
exendin-4 (a Heloderma lizard polypeptide with homology to human
GLP-1) and derivatives thereof, GLP-1 analogs modified to be DPP-IV
resistant, or human GLP-1 polypeptides conjugated to various
further agents, e.g., to extend the half-life. GLP-1
mimetics/analogues can include, e.g., exenatide, lixisenatide,
dulaglutide, semaglutide, albiglutide, LY2189265, liraglutide, and
taspoglutide. Examples of such molecules and further discussion of
their manufacture and activity can be found in the art, e.g.,
Gupta. Indian J. Endocrinol Metab 17:413-421 (2013); Garber.
Diabetes Treatments 41:S279-S284 (2018); US Patent Publication
US2009/0181912; and International Patent Publication WO2011/080103,
each of which is incorporated by reference herein in its
entirety.
[0156] In some embodiments of any of the aspects, the active
compound can be a chemotherapeutic agent or agent effective for the
treatment of cancer. As used herein, the term "cancer" relates
generally to a class of diseases or conditions in which abnormal
cells divide without control and can invade nearby tissues. Cancer
cells can also spread to other parts of the body through the blood
and lymph systems. There are several main types of cancer.
Carcinoma is a cancer that begins in the skin or in tissues that
line or cover internal organs. Sarcoma is a cancer that begins in
bone, cartilage, fat, muscle, blood vessels, or other connective or
supportive tissue. Leukemia is a cancer that starts in
blood-forming tissue such as the bone marrow, and causes large
numbers of abnormal blood cells to be produced and enter the blood.
Lymphoma and multiple myeloma are cancers that begin in the cells
of the immune system. Central nervous system cancers are cancers
that begin in the tissues of the brain and spinal cord.
[0157] In some embodiments of any of the aspects, the cancer is a
primary cancer. In some embodiments of any of the aspects, the
cancer is a malignant cancer. As used herein, the term "malignant"
refers to a cancer in which a group of tumor cells display one or
more of uncontrolled growth (i.e., division beyond normal limits),
invasion (i.e., intrusion on and destruction of adjacent tissues),
and metastasis (i.e., spread to other locations in the body via
lymph or blood). As used herein, the term "metastasize" refers to
the spread of cancer from one part of the body to another. A tumor
formed by cells that have spread is called a "metastatic tumor" or
a "metastasis." The metastatic tumor contains cells that are like
those in the original (primary) tumor. As used herein, the term
"benign" or "non-malignant" refers to tumors that may grow larger
but do not spread to other parts of the body. Benign tumors are
self-limited and typically do not invade or metastasize.
[0158] A "cancer cell" or "tumor cell" refers to an individual cell
of a cancerous growth or tissue. A tumor refers generally to a
swelling or lesion formed by an abnormal growth of cells, which may
be benign, pre-malignant, or malignant. Most cancer cells form
tumors, but some, e.g., leukemia, do not necessarily form tumors.
For those cancer cells that form tumors, the terms cancer (cell)
and tumor (cell) are used interchangeably.
[0159] As used herein the term "neoplasm" refers to any new and
abnormal growth of tissue, e.g., an abnormal mass of tissue, the
growth of which exceeds and is uncoordinated with that of the
normal tissues. Thus, a neoplasm can be a benign neoplasm,
premalignant neoplasm, or a malignant neoplasm.
[0160] A subject that has a cancer or a tumor is a subject having
objectively measurable cancer cells present in the subject's body.
Included in this definition are malignant, actively proliferative
cancers, as well as potentially dormant tumors or micrometastases.
Cancers which migrate from their original location and seed other
vital organs can eventually lead to the death of the subject
through the functional deterioration of the affected organs.
[0161] Examples of cancer include but are not limited to,
carcinoma, lymphoma, blastoma, sarcoma, leukemia, basal cell
carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain
and CNS cancer; breast cancer; cancer of the peritoneum; cervical
cancer; choriocarcinoma; colon and rectum cancer; connective tissue
cancer; cancer of the digestive system; endometrial cancer;
esophageal cancer; eye cancer; cancer of the head and neck; gastric
cancer (including gastrointestinal cancer); glioblastoma (GBM);
hepatic carcinoma; hepatoma; intra-epithelial neoplasm; kidney or
renal cancer; larynx cancer; leukemia; liver cancer; lung cancer
(e.g., small-cell lung cancer, non-small cell lung cancer,
adenocarcinoma of the lung, and squamous carcinoma of the lung);
lymphoma including Hodgkin's and non-Hodgkin's lymphoma; melanoma;
myeloma; neuroblastoma; oral cavity cancer (e.g., lip, tongue,
mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate
cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of
the respiratory system; salivary gland carcinoma; sarcoma; skin
cancer; squamous cell cancer; stomach cancer; testicular cancer;
thyroid cancer; uterine or endometrial cancer; cancer of the
urinary system; vulval cancer; as well as other carcinomas and
sarcomas; as well as B-cell lymphoma (including low
grade/follicular non-Hodgkin's lymphoma (NHL); small lymphocytic
(SL) NHL; intermediate grade/follicular NHL; intermediate grade
diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic
NHL; high grade small non-cleaved cell NHL; bulky disease NHL;
mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom's
Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute
lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic
myeloblastic leukemia; and post-transplant lymphoproliferative
disorder (PTLD), as well as abnormal vascular proliferation
associated with phakomatoses, edema (such as that associated with
brain tumors), and Meigs' syndrome.
[0162] A "cancer cell" is a cancerous, pre-cancerous, or
transformed cell, either in vivo, ex vivo, or in tissue culture,
that has spontaneous or induced phenotypic changes that do not
necessarily involve the uptake of new genetic material. Although
transformation can arise from infection with a transforming virus
and incorporation of new genomic nucleic acid, or uptake of
exogenous nucleic acid, it can also arise spontaneously or
following exposure to a carcinogen, thereby mutating an endogenous
gene. Transformation/cancer is associated with, e.g., morphological
changes, immortalization of cells, aberrant growth control, foci
formation, anchorage independence, malignancy, loss of contact
inhibition and density limitation of growth, growth factor or serum
independence, tumor specific markers, invasiveness or metastasis,
and tumor growth in suitable animal hosts such as nude mice.
[0163] In some embodiments of any of the aspects, the composition
as described herein, e.g., a composition comprising at least one IL
as described herein in combination with at least one active
compound, is administered as a monotherapy, e.g., another treatment
for the condition is not administered to the subject.
[0164] In some embodiments of any of the aspects, the methods
described herein can further comprise administering a second agent
and/or treatment to the subject, e.g. as part of a combinatorial
therapy, either in the composition described herein, e.g., a
composition comprising at least one IL as described herein in
combination with at least one active compound, or as a separate
formulation. For example, non-limiting examples of a second agent
and/or treatment for treatment of cancer can include radiation
therapy, surgery, gemcitabine, cisplastin, paclitaxel, carboplatin,
bortezomib, AMG479, vorinostat, rituximab, temozolomide, rapamycin,
ABT-737, PI-103; alkylating agents such as thiotepa and
CYTOXAN.RTM. cyclosphosphamide; alkyl sulfonates such as busulfan,
improsulfan and piposulfan; aziridines such as benzodopa,
carboquone, meturedopa, and uredopa; ethylenimines and
methylamelamines including altretamine, triethylenemelamine,
trietylenephosphoramide, triethiylenethiophosphoramide and
trimethylolomelamine; acetogenins (especially bullatacin and
bullatacinone); a camptothecin (including the synthetic analogue
topotecan); bryostatin; callystatin; CC-1065 (including its
adozelesin, carzelesin and bizelesin synthetic analogues);
cryptophycins (particularly cryptophycin 1 and cryptophycin 8);
dolastatin; duocarmycin (including the synthetic analogues, KW-2189
and CB1-TM1); eleutherobin; pancratistatin; a sarcodictyin;
spongistatin; nitrogen mustards such as chlorambucil,
chlornaphazine, cholophosphamide, estramustine, ifosfamide,
mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,
novembichin, phenesterine, prednimustine, trofosfamide, uracil
mustard; nitrosureas such as carmustine, chlorozotocin,
fotemustine, lomustine, nimustine, and ranimnustine; antibiotics
such as the enediyne antibiotics (e.g., calicheamicin, especially
calicheamicin gamma1I and calicheamicin omegaI1 (see, e.g., Agnew,
Chem. Intl. Ed. Engl., 33: 183-186 (1994)); dynemicin, including
dynemicin A; bisphosphonates, such as clodronate; an esperamicin;
as well as neocarzinostatin chromophore and related chromoprotein
enediyne antibiotic chromophores), aclacinomysins, actinomycin,
authramycin, azaserine, bleomycins, cactinomycin, carabicin,
caminomycin, carzinophilin, chromomycinis, dactinomycin,
daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine,
ADRIAMYCIN.RTM. doxorubicin (including morpholino-doxorubicin,
cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and
deoxydoxorubicin), epirubicin, esorubicin, idarubicin,
marcellomycin, mitomycins such as mitomycin C, mycophenolic acid,
nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin,
quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin,
ubenimex, zinostatin, zorubicin; anti-metabolites such as
methotrexate and 5-fluorouracil (5-FU); folic acid analogues such
as denopterin, methotrexate, pteropterin, trimetrexate; purine
analogs such as fludarabine, 6-mercaptopurine, thiamiprine,
thioguanine; pyrimidine analogs such as ancitabine, azacitidine,
6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine,
enocitabine, floxuridine; androgens such as calusterone,
dromostanolone propionate, epitiostanol, mepitiostane,
testolactone; anti-adrenals such as aminoglutethimide, mitotane,
trilostane; folic acid replenisher such as frolinic acid;
aceglatone; aldophosphamide glycoside; aminolevulinic acid;
eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate;
defofamine; demecolcine; diaziquone; elformithine; elliptinium
acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea;
lentinan; lonidainine; maytansinoids such as maytansine and
ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine;
pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic
acid; 2-ethylhydrazide; procarbazine; PSK.RTM. polysaccharide
complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin;
sizofuran; spirogermanium; tenuazonic acid; triaziquone;
2,2',2''-trichlorotriethylamine; trichothecenes (especially T-2
toxin, verracurin A, roridin A and anguidine); urethan; vindesine;
dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman;
gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa;
taxoids, e.g., TAXOL.RTM. paclitaxel (Bristol-Myers Squibb
Oncology, Princeton, N.J.), ABRAXANE.RTM. Cremophor-free,
albumin-engineered nanoparticle formulation of paclitaxel (American
Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE.RTM.
doxetaxel (Rhone-Poulenc Rorer, Antony, France); chloranbucil;
GEMZAR.RTM. gemcitabine; 6-thioguanine; mercaptopurine;
methotrexate; platinum analogs such as cisplatin, oxaliplatin and
carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide;
mitoxantrone; vincristine; NAVELBINE.RTM. vinorelbine; novantrone;
teniposide; edatrexate; daunomycin; aminopterin; xeloda;
ibandronate; irinotecan (Camptosar, CPT-11) (including the
treatment regimen of irinotecan with 5-FU and leucovorin);
topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO);
retinoids such as retinoic acid; capecitabine; combretastatin;
leucovorin (LV); oxaliplatin, including the oxaliplatin treatment
regimen (FOLFOX); lapatinib (Tykerb.RTM.); inhibitors of PKC-alpha,
Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva.RTM.)) and VEGF-A that
reduce cell proliferation and pharmaceutically acceptable salts,
acids or derivatives of any of the above. In addition, the methods
of treatment can further include the use of radiation or radiation
therapy. Further, the methods of treatment can further include the
use of surgical treatments.
[0165] Due to the excellent transdermal drug delivery
characteristics of the ILs described herein, the compositions and
combinations described herein are suitable for use with active
compounds effective in treating skin diseases and alopoeica, e.g.,
alopoecia areta. Suitable active compounds for the treatment of
alopoeica can include, e.g., corticosteroids (e.g., clobetasol or
fluocinonide), minoxidil, Elocon (mometasone), irritants (e.g.,
anthralin or topical coal tar), and ciclosporin.
[0166] In certain embodiments, an effective dose of a composition
described herein, e.g, a composition comprising at least one IL as
described herein in combination with at least one active compound,
can be administered to a patient once. In certain embodiments, an
effective dose a composition described herein, e.g, a composition
comprising at least one IL as described herein in combination with
at least one active compound, can be administered to a patient
repeatedly. For systemic administration, subjects can be
administered a therapeutic amount of a composition described
herein, e.g, a composition comprising at least one IL as described
herein in combination with at least one active compound, such as,
e.g. 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5
mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg,
50 mg/kg, or more. In some embodiments of any of the aspects, the
at least one active compound is present in the combination at a
dose of from about 1.0-20.0 mg/kg. In some embodiments of any of
the aspects, the at least one active compound is present in the
combination at a dose of from 1.0-20.0 mg/kg.
[0167] In some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be from about 1 U/kg to
about 20 U/kg. In some embodiments, the active compound is insulin
and the concentration or dosage of insulin can be from 1 U/kg to 20
U/kg. In some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be less than 20 U/kg. In
some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be from about 2 U/kg to
about 10 U/kg. In some embodiments, the active compound is insulin
and the concentration or dosage of insulin can be from 2 U/kg to 10
U/kg. In some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be from about 2 U/kg to
about 5 U/kg. In some embodiments, the active compound is insulin
and the concentration or dosage of insulin can be from 2 U/kg to 5
U/kg. In some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be from about 5 U/kg to
about 10 U/kg. In some embodiments, the active compound is insulin
and the concentration or dosage of insulin can be from 5 U/kg to 10
U/kg. In some embodiments, the active compound is insulin and the
concentration or dosage of insulin can be 2 U/kg, 5 U/kg, or 10
U/kg.
[0168] In one aspect of any of the embodiments, described herein is
a method of treating a disease in a subject in need thereof by
administering to the subject an active compound in combination with
the at least one IL as described herein by into the affected tissue
by injection. In some embodiments, the affected tissue is tissue
comprising diseased cells. In some embodiments, the affected tissue
is tissue displaying symptoms of the disease. Non-limiting examples
of suitable affected tissues include tumor tissue, fat tissue,
adipose tissue, or the like. In some embodiments of any of the
aspects, suitable affected tissues include tumor tissue, fat
tissue, adipose tissue, or the like. In some embodiments of any of
the aspects, the disease is a disease arising from tissue growth,
e.g., unwanted, aberrant, or pathological tissue growth. A disease
arising from tissue growth can be any disease caused by or
characterized by, a rate of tissue growth, location of tissue
growth, or pattern/structure of tissue growth which differs from
what is normal for that tissue type in a healthy subject.
Non-limiting examples of such diseases are tumors, cancer,
fat/obesity, and/or hyperplasia. In some embodiments of any of the
aspects, such diseases are tumors, cancer, fat/obesity, and/or
hyperplasia.
[0169] Enzyme inhibitors are a treatment option for a number of
conditions, including diabetes, where, for example,
insulin-degrading enzyme inhibitors, ACE inhibitors, and
alpha-glucosidase inhibitors have all been explored as therapeutic
approaches. Safe, effective enzyme inhibitors are therefore of
interest in the treatment of a number of conditions. Without
wishing to be bound by theory, it is contemplated herein that the
ILs described herein can exhibit enzyme inhibition activity.
Accordingly, in one aspect of any of the embodiments, described
herein is a method of treating diabetes, ulcers, cancer, or
fibrosis in a subject in need thereof, the method comprising
administering to the subject a composition comprising at least one
IL as described herein. In some embodiments, the composition does
not comprise a further therapeutically active agent.
[0170] Fibrotic conditions benefit from the production and/or
maintenance of the extracellular matrix by reducing the
accumulation of scar tissue in favor of extracellular matrix. As
used herein, "fibrosis" refers to the formation of fibrous tissue
as a reparative or reactive process, rather than as a normal
constituent of an organ or tissue. Fibrosis is characterized by
fibroblast accumulation and collagen deposition in excess of normal
deposition in any particular tissue. Fibrosis can occur as the
result of inflammation, irritation, or healing. A subject in need
of treatment for a fibrotic condition is any subject having, or
diagnosed as having, or at risk of having a fibrotic condition.
Non-limiting examples of fibrotic conditions include, but are not
limited to pulmonary fibrosis; scarring; scarring of the skin;
trauma; a wound; chronic wounds (e.g. as in diabetes patients),
corneal defects; corneal ulceration; corneal wounds; diabetic
ulcer; ulcer; sepsis; arthritis; idiopathic pulmonary fibrosis;
cystic fibrosis; cirrhosis; endomyocardial fibrosis; mediastinal
fibrosis; myelofibrosis; retroperitoneal fibrosis; progressive
massive fibrosis; nephrogenic systemic fibrosis; Crohn's disease;
keloid; scleroderma; systemic sclerosis; arthrofibrosis; adhesive
capsulitis; lung fibrosis; liver fibrosis; kidney fibrosis; heart
fibrosis; vascular fibrosis; skin fibrosis; eye fibrosis; bone
marrow fibrosis; asthma; sarcoidosis; COPD; emphysema;
nschistomasomiasis; cholangitis; diabetic nephropathy; lupus
nephritis; postangioplasty arterial restenosis; atherosclerosis;
burn scarring; hypertrophic scarring; nephrogenic fibrosing
dermatopathy; postcataract surgery; proliferative
vitreoretinopathy; Peyronie's disease; Duputren's contracture;
dermatomyositis; and graft versus host disease.
[0171] As used herein, "ulcer" refers to a break or disruption of a
bodily membrane. In some embodiments, the ulcer can be caused by
inflammation and/or necrosis of the affected tissue. Ulcers can be
skin ulcers (e.g., pressure ulcers, diabetic ulcers, ulcerative
dermatitis, and the like), a corneal ulcer, an oral ulcer, a peptic
ulcer, a venousucler, a stress ulcer, or ulcerative colitis.
[0172] In some embodiments, after an initial treatment regimen, the
treatments can be administered on a less frequent basis. For
example, after treatment biweekly for three months, treatment can
be repeated once per month, for six months or a year or longer.
Treatment according to the methods described herein can reduce
levels of a marker or symptom of a condition, by at least 10%, at
least 15%, at least 20%, at least 25%, at least 30%, at least 40%,
at least 50%, at least 60%, at least 70%, at least 80% or at least
90% or more.
[0173] The dosage of a composition as described herein can be
determined by a physician and adjusted, as necessary, to suit
observed effects of the treatment. With respect to duration and
frequency of treatment, it is typical for skilled clinicians to
monitor subjects in order to determine when the treatment is
providing therapeutic benefit, and to determine whether to increase
or decrease dosage, increase or decrease administration frequency,
discontinue treatment, resume treatment, or make other alterations
to the treatment regimen. The dosing schedule can vary from once a
week to daily depending on a number of clinical factors, such as
the subject's sensitivity to the active compound. The desired dose
or amount of activation can be administered at one time or divided
into subdoses, e.g., 2-4 subdoses and administered over a period of
time, e.g., at appropriate intervals through the day or other
appropriate schedule. In some embodiments, administration can be
chronic, e.g., one or more doses and/or treatments daily over a
period of weeks or months. Examples of dosing and/or treatment
schedules are administration daily, twice daily, three times daily
or four or more times daily over a period of 1 week, 2 weeks, 3
weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or
6 months, or more. A composition described herein, e.g, a
composition comprising at least one IL in combination with at least
one active compound, can be administered over a period of time,
such as over a 5 minute, 10 minute, 15 minute, 20 minute, or 25
minute period.
[0174] The dosage ranges for the administration of the compositions
described herein, according to the methods described herein depend
upon, for example, the form of the active compound, its potency,
and the extent to which symptoms, markers, or indicators of a
condition described herein are desired to be reduced, for example
the percentage reduction desired for symptoms or markers. The
dosage should not be so large as to cause adverse side effects.
Generally, the dosage will vary with the age, condition, and sex of
the patient and can be determined by one of skill in the art. The
dosage can also be adjusted by the individual physician in the
event of any complication.
[0175] The efficacy of a composition described in, e.g. the
treatment of a condition described herein, or to induce a response
as described herein can be determined by the skilled clinician.
However, a treatment is considered "effective treatment," as the
term is used herein, if one or more of the signs or symptoms of a
condition described herein are altered in a beneficial manner,
other clinically accepted symptoms are improved, or even
ameliorated, or a desired response is induced e.g., by at least 10%
following treatment according to the methods described herein.
Efficacy can be assessed, for example, by measuring a marker,
indicator, symptom, and/or the incidence of a condition treated
according to the methods described herein or any other measurable
parameter appropriate. Efficacy can also be measured by a failure
of an individual to worsen as assessed by hospitalization, or need
for medical interventions (i.e., progression of the disease is
halted). Methods of measuring these indicators are known to those
of skill in the art and/or are described herein. Treatment includes
any treatment of a disease in an individual or an animal (some
non-limiting examples include a human or an animal) and includes:
(1) inhibiting the disease, e.g., preventing a worsening of
symptoms (e.g. pain or inflammation); or (2) relieving the severity
of the disease, e.g., causing regression of symptoms. An effective
amount for the treatment of a disease means that amount which, when
administered to a subject in need thereof, is sufficient to result
in effective treatment as that term is defined herein, for that
disease. Efficacy of an agent can be determined by assessing
physical indicators of a condition or desired response. It is well
within the ability of one skilled in the art to monitor efficacy of
administration and/or treatment by measuring any one of such
parameters, or any combination of parameters. Efficacy can be
assessed in animal models of a condition described herein, for
example treatment of diabetes or cancer. When using an experimental
animal model, efficacy of treatment is evidenced when a
statistically significant change in a marker is observed.
[0176] In vitro and animal model assays are provided herein which
allow the assessment of a given dose of a composition described
herein, e.g, a composition comprising at least one IL in
combination with at least one active compound. By way of
non-limiting example, the effects of a dose of a composition
comprising at least one IL in combination with insulin can be
assessed by using the models described in the Examples herein.
[0177] The incidence of obesity is on the rise and existing
treatments, such as diets, have notoriously low long-term success
rates. Additional treatments and strategies for reducing obesity or
reducing weight gain rates are of critical importance both for
addressing obesity itself as well the number of conditions that are
caused by or exacerbated by excess weight. Without wishing to be
bound by theory, it is contemplated herein that the ILs as
described herein can reduce the uptake of hydrophobic/lipophilic
molecules in the intestine. Accordingly, provided herein are
methods of treating obesity and/or reducing weight/weight gain by
administering at least one IL as described herein to a subject in
need of such. In some embodiments of any of the aspects, a subject
treated in accordance with the present methods is a subject not
having or not diagnosed as having diabetes. In some embodiments of
any of the aspects, a subject treated in accordance with the
present methods is a subject not administered insulin. In some
embodiments of any of the aspects, the composition comprising at
least one IL does not comprise insulin. In some embodiments of any
of the aspects, the composition comprising at least one IL does not
comprise another pharmaceutically active ingredient and/or another
agent which is therapeutically effective in treating diabetes.
[0178] In some embodiments of any of the aspects, the active
compound is therapeutically effective in treating obesity. In some
embodiments of any of the aspects, the active compound is
therapeutically effective in treating a disease associated with
obesity. In some embodiments of any of the aspects, the active
compound is therapeutically effective in treating a disease caused
by obesity. In some embodiments of any of the aspects, the active
compound is therapeutically effective in treating a disease which
causes obesity. In some embodiments of any of the aspects, the
active compound is therapeutically effective in treating metabolic
syndrome.
[0179] In some embodiments of any of the aspects, the subject
administered a composition comprising at least one IL as described
herein, e.g., in combination with an active compound is a subject
having, diagnosed as having, or in need of treatment for obesity,
excess weight, or prevention of weight gain. In some embodiments,
the subject is overweight. The methods described herein comprises
methods of treating obesity, reducing weight gain, preventing
weight gain, promoting weight loss, and the like. Such methods can,
e.g., promote metabolic health, be pursued for aesthetic reasons,
and/or prepare patients for surgical interventions which are
counterindicated for those with high BMIs or weights. In some
embodiments, weight loss can be medically necessary and/or
medically indicated, e.g. when the subject is overweight and/or
obese. In some embodiments, weight loss can be for cosmetic
purposes, e.g. when the subject desires to lose weight whether or
not weight loss is medically necessary and/or medically
indicated.
[0180] The term "obesity" refers to excess fat in the body. Obesity
can be determined by any measure accepted and utilized by those of
skill in the art. Currently, an accepted measure of obesity is body
mass index (BMI), which is a measure of body weight in kilograms
relative to the square of height in meters. Generally, for an adult
over age 20, a BMI between about 18.5 and 24.9 is considered
normal, a BMI between about 25.0 and 29.9 is considered overweight,
a BMI at or above about 30.0 is considered obese, and a BMI at or
above about 40 is considered morbidly obese. (See, e.g., Gallagher
et al. (2000) Am J Clin Nutr 72:694-701.) These BMI ranges are
based on the effect of body weight on increased risk for disease.
Some common conditions related to high BMI and obesity include
cardiovascular disease, high blood pressure (i.e., hypertension),
osteoarthritis, cancer, and diabetes. Although BMI correlates with
body fat, the relation between BMI and actual body fat differs with
age and gender. For example, women are more likely to have a higher
percent of body fat than men for the same BMI. Furthermore, the BMI
threshold that separates normal, overweight, and obese can vary,
e.g. with age, gender, ethnicity, fitness, and body type, amongst
other factors. In some embodiments, a subject with obesity can be a
subject with a body mass index of at least about 25 kg/m.sup.2
prior to administration of a treatment as described herein. In some
embodiments, a subject with obesity can be a subject with a body
mass index of at least about 30 kg/m.sup.2 prior to administration
of a treatment as described herein.
[0181] In some embodiments of any of the aspects, the subject
administered a composition comprising at least one IL as described
herein, e.g., in combination with at least one active compound is a
subject having, diagnosed as having, or in need of treatment for a
metabolic disorder or metabolic syndrome. The term "metabolic
disorder" refers to any disorder associated with or aggravated by
impaired or altered glucose regulation or glycemic control, such
as, for example, insulin resistance. Such disorders include, but
are not limited to obesity; excess adipose tissue; diabetes; fatty
liver disease; non-alcoholic fatty liver disease; metabolic
syndrome; dyslipidemia; hypertension; hyperglycemia; and
cardiovascular disease. "Metabolic syndrome", which is distinct
from metabolic disorder, refers to a combination of medical
disorders that, when occurring together, increase the risk of
developing cardiovascular disease and diabetes. A number of
definitions of metabolic syndrome have been established, e.g by the
American Heart Association and the International Diabetes
Foundation. As but one example, the WHO defines metabolic syndrome
as the presence of any one of diabetes mellitus, impaired glucose
tolerance, impaired fasting glucose or insulin resistance and two
of the following: blood pressure equal to or greater than 140/90
mmHg, dyslipidemia, central obesity, and microalbuminuria. In some
embodiments, the metabolic disorder can be selected from the group
consisting of: obesity; excess adipose tissue; diabetes; and
cardiovascular disease.
[0182] The uptake of many active compounds, e.g., pharmaceutically
active compounds, can be improved by delivering the compounds in
solvents. However, such approaches are often unsuitable for in vivo
use because most such solvents demonstrate toxic side effects
and/or act as irritants to the point of delivery. Described herein
are methods and compositions which can provide low toxicity with
improved delivery kinetics.
[0183] For convenience, the meaning of some terms and phrases used
in the specification, examples, and appended claims, are provided
below. Unless stated otherwise, or implicit from context, the
following terms and phrases include the meanings provided below.
The definitions are provided to aid in describing particular
embodiments, and are not intended to limit the claimed invention,
because the scope of the invention is limited only by the claims.
Unless otherwise defined, all technical and scientific terms used
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which this invention belongs. If there
is an apparent discrepancy between the usage of a term in the art
and its definition provided herein, the definition provided within
the specification shall prevail.
[0184] For convenience, certain terms employed herein, in the
specification, examples and appended claims are collected here.
[0185] A carboxylic acid is a carbonyl-bearing functional group
having a formula RCOOH where R is aliphatic, heteroaliphatic,
alkyl, or heteroalkyl.
[0186] As used herein, the term "alkyl" means a straight or
branched, saturated aliphatic radical having a chain of carbon
atoms. The term "alkyl" includes cycloalkyl or cyclic alkyl.
C.sub.x alkyl and C.sub.x-C.sub.yalkyl are typically used where X
and Y indicate the number of carbon atoms in the chain. For
example, C.sub.1-C.sub.6alkyl includes alkyls that have a chain of
between 1 and 6 carbons (e.g., methyl, ethyl, propyl, isopropyl,
butyl, sec-butyl, isobutyl, tert-butyl, pentyl, neopentyl, hexyl,
and the like). Alkyl represented along with another radical (e.g.,
as in arylalkyl) means a straight or branched, saturated alkyl
divalent radical having the number of atoms indicated or when no
atoms are indicated means a bond, e.g.,
(C.sub.6-C.sub.10)aryl(C.sub.0-C.sub.3)alkyl includes phenyl,
benzyl, phenethyl, 1-phenylethyl 3-phenylpropyl, and the like.
Backbone of the alkyl can be optionally inserted with one or more
heteroatoms, such as N, O, or S. Examples of alkyl radicals
include, but are not limited to, methyl, ethyl, propyl, isopropyl,
n-butyl, tert-butyl, neopentyl, n-hexyl, and n-octyl radicals.
[0187] In preferred embodiments, a straight chain or branched chain
alkyl has 30 or fewer carbon atoms in its backbone (e.g.,
C.sub.1-C.sub.30 for straight chains, C.sub.3-C.sub.30 for branched
chains), and more preferably 20 or fewer. Likewise, preferred
cycloalkyls have from 3-10 carbon atoms in their ring structure,
and more preferably have 5, 6 or 7 carbons in the ring structure.
The term "alkyl" (or "lower alkyl") as used throughout the
specification, examples, and claims is intended to include both
"unsubstituted alkyls" and "substituted alkyls", the latter of
which refers to alkyl moieties having one or more substituents
replacing a hydrogen on one or more carbons of the hydrocarbon
backbone.
[0188] Unless the number of carbons is otherwise specified, "lower
alkyl" as used herein means an alkyl group, as defined above, but
having from one to ten carbons, more preferably from one to six
carbon atoms in its backbone structure. Likewise, "lower alkenyl"
and "lower alkynyl" have similar chain lengths. Throughout the
application, preferred alkyl groups are lower alkyls. In preferred
embodiments, a substituent designated herein as alkyl is a lower
alkyl.
[0189] Substituents of a substituted alkyl can include halogen,
hydroxy, nitro, thiols, amino, azido, imino, amido, phosphoryl
(including phosphonate and phosphinate), sulfonyl (including
sulfate, sulfonamido, sulfamoyl and sulfonate), and silyl groups,
as well as ethers, alkylthios, carbonyls (including ketones,
aldehydes, carboxylates, and esters), --CF3, --CN and the like.
[0190] As used herein, the term "alkenyl" refers to unsaturated
straight-chain, branched-chain or cyclic hydrocarbon radicals
having at least one carbon-carbon double bond. C.sub.x alkenyl and
C.sub.x-C.sub.yalkenyl are typically used where X and Y indicate
the number of carbon atoms in the chain. For example,
C.sub.2-C.sub.6alkenyl includes alkenyls that have a chain of
between 1 and 6 carbons and at least one double bond, e.g., vinyl,
allyl, propenyl, isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl,
2-methylallyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, and the like).
Alkenyl represented along with another radical (e.g., as in
arylalkenyl) means a straight or branched, alkenyl divalent radical
having the number of atoms indicated. Backbone of the alkenyl can
be optionally inserted with one or more heteroatoms, such as N, O,
or S.
[0191] As used herein, the term "alkynyl" refers to unsaturated
hydrocarbon radicals having at least one carbon-carbon triple bond.
C.sub.x alkynyl and C.sub.x-C.sub.yalkynyl are typically used where
X and Y indicate the number of carbon atoms in the chain. For
example, C.sub.2-C.sub.6alkynyl includes alkynls that have a chain
of between 1 and 6 carbons and at least one triple bond, e.g.,
ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, isopentynyl,
1,3-hexa-diyn-yl, n-hexynyl, 3-pentynyl, 1-hexen-3-ynyl and the
like. Alkynyl represented along with another radical (e.g., as in
arylalkynyl) means a straight or branched, alkynyl divalent radical
having the number of atoms indicated. Backbone of the alkynyl can
be optionally inserted with one or more heteroatoms, such as N, O,
or S.
[0192] As used herein, the term "halogen" or "halo" refers to an
atom selected from fluorine, chlorine, bromine and iodine. The term
"halogen radioisotope" or "halo isotope" refers to a radionuclide
of an atom selected from fluorine, chlorine, bromine and iodine. A
"halogen-substituted moiety" or "halo-substituted moiety", as an
isolated group or part of a larger group, means an aliphatic,
alicyclic, or aromatic moiety, as described herein, substituted by
one or more "halo" atoms, as such terms are defined in this
application. For example, halo-substituted alkyl includes
haloalkyl, dihaloalkyl, trihaloalkyl, perhaloalkyl and the like
(e.g. halosubstituted (C.sub.1-C.sub.3)alkyl includes chloromethyl,
dichloromethyl, difluoromethyl, trifluoromethyl (--CF.sub.3),
2,2,2-trifluoroethyl, perfluoroethyl,
2,2,2-trifluoro-1,1-dichloroethyl, and the like).
[0193] The term "cyclyl" or "cycloalkyl" refers to saturated and
partially unsaturated cyclic hydrocarbon groups having 3 to 12
carbons, for example, 3 to 8 carbons, and, for example, 3 to 6
carbons. C.sub.xcyclyl and C.sub.x-C.sub.ycyclyl are typically used
where X and Y indicate the number of carbon atoms in the ring
system. The cycloalkyl group additionally can be optionally
substituted, e.g., with 1, 2, 3, or 4 substituents. Examples of
cyclyl groups include, without limitation, cyclopropyl, cyclobutyl,
cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,
2,5-cyclohexadienyl, cycloheptyl, cyclooctyl, bicyclo[2.2.2]octyl,
adamantan-1-yl, decahydronaphthyl, oxocyclohexyl, dioxocyclohexyl,
thiocyclohexyl, 2-oxobicyclo [2.2.1]hept-1-yl, and the like
[0194] The term "heterocyclyl" refers to a nonaromatic 5-8 membered
monocyclic, 8-12 membered bicyclic, or 11-14 membered tricyclic
ring system having 1-3 heteroatoms if monocyclic, 1-6 heteroatoms
if bicyclic, or 1-9 heteroatoms if tricyclic, said heteroatoms
selected from O, N, or S (e.g., carbon atoms and 1-3, 1-6, or 1-9
heteroatoms of N, O, or S if monocyclic, bicyclic, or tricyclic,
respectively). C.sub.xheterocyclyl and C.sub.x-C.sub.yheterocyclyl
are typically used where X and Y indicate the number of carbon
atoms in the ring system. In some embodiments, 1, 2 or 3 hydrogen
atoms of each ring can be substituted by a substituent. Exemplary
heterocyclyl groups include, but are not limited to piperazinyl,
pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, piperidyl,
4-morpholyl, 4-piperazinyl, pyrrolidinyl, perhydropyrrolizinyl,
1,4-diazaperhydroepinyl, 1,3-dioxanyl, 1,4-dioxanyland the
like.
[0195] The terms "bicyclic" and "tricyclic" refers to fused,
bridged, or joined by a single bond polycyclic ring assemblies. As
used herein, the term "fused ring" refers to a ring that is bonded
to another ring to form a compound having a bicyclic structure when
the ring atoms that are common to both rings are directly bound to
each other. Non-exclusive examples of common fused rings include
decalin, naphthalene, anthracene, phenanthrene, indole, furan,
benzofuran, quinoline, and the like. Compounds having fused ring
systems can be saturated, partially saturated, cyclyl,
heterocyclyl, aromatics, heteroaromatics, and the like.
[0196] The term "aryl" refers to monocyclic, bicyclic, or tricyclic
fused aromatic ring system. C.sub.x aryl and C.sub.x-C.sub.yaryl
are typically used where X and Y indicate the number of carbon
atoms in the ring system. Exemplary aryl groups include, but are
not limited to, pyridinyl, pyrimidinyl, furanyl, thienyl,
imidazolyl, thiazolyl, pyrazolyl, pyridazinyl, pyrazinyl,
triazinyl, tetrazolyl, indolyl, benzyl, phenyl, naphthyl,
anthracenyl, azulenyl, fluorenyl, indanyl, indenyl, naphthyl,
phenyl, tetrahydronaphthyl, benzimidazolyl, benzofuranyl,
benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl,
benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl,
benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl,
carbolinyl, chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl,
2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3 b]tetrahydrofuran,
furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl,
1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl,
3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl,
isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl,
methylenedioxyphenyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,
oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl,
phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl,
phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl,
4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl,
pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole,
pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl,
pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl,
quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl,
tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl and xanthenyl, and the like. In some
embodiments, 1, 2, 3, or 4 hydrogen atoms of each ring can be
substituted by a substituent.
[0197] The term "heteroaryl" refers to an aromatic 5-8 membered
monocyclic, 8-12 membered fused bicyclic, or 11-14 membered fused
tricyclic ring system having 1-3 heteroatoms if monocyclic, 1-6
heteroatoms if bicyclic, or 1-9 heteroatoms if tricyclic, said
heteroatoms selected from O, N, or S (e.g., carbon atoms and 1-3,
1-6, or 1-9 heteroatoms of N, O, or S if monocyclic, bicyclic, or
tricyclic, respectively. C.sub.x heteroaryl and
C.sub.x-C.sub.yheteroaryl are typically used where X and Y indicate
the number of carbon atoms in the ring system. Heteroaryls include,
but are not limited to, those derived from benzo[b]furan, benzo[b]
thiophene, benzimidazole, imidazo[4,5-c]pyridine, quinazoline,
thieno[2,3-c]pyridine, thieno[3,2-b]pyridine,
thieno[2,3-b]pyridine, indolizine, imidazo[1,2a]pyridine,
quinoline, isoquinoline, phthalazine, quinoxaline, naphthyridine,
quinolizine, indole, isoindole, indazole, indoline, benzoxazole,
benzopyrazole, benzothiazole, imidazo[1,5-a]pyridine,
pyrazolo[1,5-a]pyridine, imidazo[1,2-a]pyrimidine,
imidazo[1,2-c]pyrimidine, imidazo[1,5-a]pyrimidine,
imidazo[1,5-c]pyrimidine, pyrrolo[2,3-b]pyridine,
pyrrolo[2,3cjpyridine, pyrrolo[3,2-c]pyridine,
pyrrolo[3,2-b]pyridine, pyrrolo[2,3-d]pyrimidine,
pyrrolo[3,2-d]pyrimidine, pyrrolo[2,3-b]pyrazine,
pyrazolo[1,5-a]pyridine, pyrrolo[1,2-b]pyridazine,
pyrrolo[1,2-c]pyrimidine, pyrrolo[1,2-a]pyrimidine,
pyrrolo[1,2-a]pyrazine, triazo[1,5-a]pyridine, pteridine, purine,
carbazole, acridine, phenazine, phenothiazene, phenoxazine,
1,2-dihydropyrrolo[3,2,1-hi]indole, indolizine,
pyrido[1,2-a]indole, 2(1H)-pyridinone, benzimidazolyl,
benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl,
benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl,
benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl,
4aH-carbazolyl, carbolinyl, chromanyl, chromenyl, cinnolinyl,
decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl,
dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl,
imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl,
indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl,
isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl,
isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl,
methylenedioxyphenyl, morpholinyl, naphthyridinyl,
octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl,
1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl,
oxazolidinyl, oxazolyl, oxepanyl, oxetanyl, oxindolyl, pyrimidinyl,
phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl,
phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl,
piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl,
purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl,
pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole,
pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl,
2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl,
quinoxalinyl, quinuclidinyl, tetrahydrofuranyl,
tetrahydroisoquinolinyl, tetrahydropyranyl, tetrahydroquinolinyl,
tetrazolyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl,
1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl,
thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienooxazolyl,
thienoimidazolyl, thiophenyl and xanthenyl. Some exemplary
heteroaryl groups include, but are not limited to, pyridyl, furyl
or furanyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or
thienyl, pyridazinyl, pyrazinyl, quinolinyl, indolyl, thiazolyl,
naphthyridinyl, 2-amino-4-oxo-3,4-dihydropteridin-6-yl,
tetrahydroisoquinolinyl, and the like. In some embodiments, 1, 2,
3, or 4 hydrogen atoms of each ring may be substituted by a
substituent.
[0198] As used herein, the term "substituted" refers to independent
replacement of one or more of the hydrogen atoms on the substituted
moiety with substituents independently selected from, but not
limited to, alkyl, alkenyl, heterocycloalkyl, alkoxy, aryloxy,
hydroxy, amino, amido, alkylamino, arylamino, cyano, halo,
mercapto, nitro, carbonyl, acyl, aryl and heteroaryl groups.
[0199] As used herein, the term "substituted" refers to independent
replacement of one or more (typically 1, 2, 3, 4, or 5) of the
hydrogen atoms on the substituted moiety with substituents
independently selected from the group of substituents listed below
in the definition for "substituents" or otherwise specified. In
general, a non-hydrogen substituent can be any substituent that can
be bound to an atom of the given moiety that is specified to be
substituted. Examples of substituents include, but are not limited
to, acyl, acylamino, acyloxy, aldehyde, alicyclic, aliphatic,
alkanesulfonamido, alkanesulfonyl, alkaryl, alkenyl, alkoxy,
alkoxycarbonyl, alkyl, alkylamino, alkylcarbanoyl, alkylene,
alkylidene, alkylthios, alkynyl, amide, amido, amino, amino,
aminoalkyl, aralkyl, aralkylsulfonamido, arenesulfonamido,
arenesulfonyl, aromatic, aryl, arylamino, arylcarbanoyl, aryloxy,
azido, carbamoyl, carbonyl, carbonyls (including ketones, carboxy,
carboxylates, CF.sub.3, cyano (CN), cycloalkyl, cycloalkylene,
ester, ether, haloalkyl, halogen, halogen, heteroaryl,
heterocyclyl, hydroxy, hydroxy, hydroxyalkyl, imino, iminoketone,
ketone, mercapto, nitro, oxaalkyl, oxo, oxoalkyl, phosphoryl
(including phosphonate and phosphinate), silyl groups, sulfonamido,
sulfonyl (including sulfate, sulfamoyl and sulfonate), thiols, and
ureido moieties, each of which may optionally also be substituted
or unsubstituted. In some cases, two substituents, together with
the carbon(s) to which they are attached to, can form a ring.
[0200] Aryl and heteroaryls can be optionally substituted with one
or more substituents at one or more positions, for example,
halogen, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl,
amino, nitro, sulfhydryl, imino, amido, phosphate, phosphonate,
phosphinate, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl,
ketone, aldehyde, ester, a heterocyclyl, an aromatic or
heteroaromatic moiety, --CF3, --CN, or the like.
[0201] The terms "alkoxyl" or "alkoxy" as used herein refers to an
alkyl group, as defined above, having an oxygen radical attached
thereto. Representative alkoxyl groups include methoxy, ethoxy,
propyloxy, tert-butoxy, n-propyloxy, iso-propyloxy, n-butyloxy,
iso-butyloxy, and the like. An "ether" is two hydrocarbons
covalently linked by an oxygen. Accordingly, the substituent of an
alkyl that renders that alkyl an ether is or resembles an alkoxyl,
such as can be represented by one of --O-- alkyl, --O-alkenyl, and
--O-alkynyl. Aroxy can be represented by --O-aryl or O-heteroaryl,
wherein aryl and heteroaryl are as defined below. The alkoxy and
aroxy groups can be substituted as described above for alkyl.
[0202] The term "aralkyl", as used herein, refers to an alkyl group
substituted with an aryl group (e.g., an aromatic or heteroaromatic
group).
[0203] The term "alkylthio" refers to an alkyl group, as defined
above, having a sulfur radical attached thereto. In preferred
embodiments, the "alkylthio" moiety is represented by one of
--S-alkyl, --S-alkenyl, and --S-alkynyl. Representative alkylthio
groups include methylthio, ethylthio, and the like. The term
"alkylthio" also encompasses cycloalkyl groups, alkene and
cycloalkene groups, and alkyne groups. "Arylthio" refers to aryl or
heteroaryl groups.
[0204] The term "sulfinyl" means the radical --SO--. It is noted
that the sulfinyl radical can be further substituted with a variety
of substituents to form different sulfinyl groups including
sulfinic acids, sulfinamides, sulfinyl esters, sulfoxides, and the
like.
[0205] The term "sulfonyl" means the radical --SO.sub.2--. It is
noted that the sulfonyl radical can be further substituted with a
variety of substituents to form different sulfonyl groups including
sulfonic acids (--SO.sub.3H), sulfonamides, sulfonate esters,
sulfones, and the like.
[0206] The term "thiocarbonyl" means the radical --C(S)--. It is
noted that the thiocarbonyl radical can be further substituted with
a variety of substituents to form different thiocarbonyl groups
including thioacids, thioamides, thioesters, thioketones, and the
like.
[0207] As used herein, the term "amino" means --NH.sub.2. The term
"alkylamino" means a nitrogen moiety having at least one straight
or branched unsaturated aliphatic, cyclyl, or heterocyclyl radicals
attached to the nitrogen. For example, representative amino groups
include --NH.sub.2, --NHCH.sub.3, --N(CH.sub.3).sub.2,
--NH(C.sub.1-C.sub.10alkyl), --N(C.sub.1-C.sub.10alkyl).sub.2, and
the like. The term "alkylamino" includes "alkenylamino,"
"alkynylamino," "cyclylamino," and "heterocyclylamino." The term
"arylamino" means a nitrogen moiety having at least one aryl
radical attached to the nitrogen. For example --NHaryl, and
--N(aryl).sub.2. The term "heteroarylamino" means a nitrogen moiety
having at least one heteroaryl radical attached to the nitrogen.
For example --NHheteroaryl, and --N(heteroaryl).sub.2. Optionally,
two substituents together with the nitrogen can also form a ring.
Unless indicated otherwise, the compounds described herein
containing amino moieties can include protected derivatives
thereof. Suitable protecting groups for amino moieties include
acetyl, tertbutoxycarbonyl, benzyloxycarbonyl, and the like.
[0208] The term "aminoalkyl" means an alkyl, alkenyl, and alkynyl
as defined above, except where one or more substituted or
unsubstituted nitrogen atoms (--N--) are positioned between carbon
atoms of the alkyl, alkenyl, or alkynyl. For example, an
(C.sub.2-C.sub.6) aminoalkyl refers to a chain comprising between 2
and 6 carbons and one or more nitrogen atoms positioned between the
carbon atoms.
[0209] The term "alkoxyalkoxy" means --O-(alkyl)-O-(alkyl), such as
--OCH.sub.2CH.sub.2OCH.sub.3, and the like. The term
"alkoxycarbonyl" means --C(O)O-(alkyl), such as
--C(.dbd.O)OCH.sub.3, --C(.dbd.O)OCH.sub.2CH.sub.3, and the like.
The term "alkoxyalkyl" means -(alkyl)-O-(alkyl), such as
--CH.sub.2OCH.sub.3, --CH.sub.2OCH.sub.2CH.sub.3, and the like. The
term "aryloxy" means --O-(aryl), such as --O-phenyl, --O-pyridinyl,
and the like. The term "arylalkyl" means -(alkyl)-(aryl), such as
benzyl (i.e., --CH.sub.2phenyl), --CH.sub.2-pyrindinyl, and the
like. The term "arylalkyloxy" means --O-(alkyl)-(aryl), such as
--O-benzyl, --O--CH.sub.2-pyridinyl, and the like. The term
"cycloalkyloxy" means --O-(cycloalkyl), such as --O-cyclohexyl, and
the like. The term "cycloalkylalkyloxy" means
--O-(alkyl)-(cycloalkyl, such as --OCH.sub.2cyclohexyl, and the
like. The term "aminoalkoxy" means --O-(alkyl)-NH.sub.2, such as
--OCH.sub.2NH.sub.2, --OCH.sub.2CH.sub.2NH.sub.2, and the like. The
term "mono- or di-alkylamino" means --NH(alkyl) or
--N(alkyl)(alkyl), respectively, such as --NHCH.sub.3,
--N(CH.sub.3).sub.2, and the like. The term "mono- or
di-alkylaminoalkoxy" means --O-(alkyl)-NH(alkyl) or
--O-(alkyl)-N(alkyl)(alkyl), respectively, such as
--OCH.sub.2NHCH.sub.3, --OCH.sub.2CH.sub.2N(CH.sub.3).sub.2, and
the like. The term "arylamino" means --NH(aryl), such as
--NH-phenyl, --NH-pyridinyl, and the like. The term
"arylalkylamino" means --NH-(alkyl)-(aryl), such as --NH-benzyl,
--NHCH.sub.2-pyridinyl, and the like. The term "alkylamino" means
--NH(alkyl), such as --NHCH.sub.3, --NHCH.sub.2CH.sub.3, and the
like. The term "cycloalkylamino" means --NH-(cycloalkyl), such as
--NH-cyclohexyl, and the like. The term "cycloalkylalkylamino"
--NH-(alkyl)-(cycloalkyl), such as --NHCH.sub.2-cyclohexyl, and the
like.
[0210] It is noted in regard to all of the definitions provided
herein that the definitions should be interpreted as being open
ended in the sense that further substituents beyond those specified
may be included. Hence, a C.sub.1 alkyl indicates that there is one
carbon atom but does not indicate what are the substituents on the
carbon atom. Hence, a C.sub.1 alkyl comprises methyl (i.e.,
--CH.sub.3) as well as --CR.sub.aR.sub.bR.sub.c where R.sub.a,
R.sub.b, and R.sub.c can each independently be hydrogen or any
other substituent where the atom alpha to the carbon is a
heteroatom or cyano. Hence, CF.sub.3, CH.sub.2OH and CH.sub.2CN are
all C.sub.1 alkyls.
[0211] Unless otherwise stated, structures depicted herein are
meant to include compounds which differ only in the presence of one
or more isotopically enriched atoms. For example, compounds having
the present structure except for the replacement of a hydrogen atom
by a deuterium or tritium, or the replacement of a carbon atom by a
.sup.13C- or .sup.14C-enriched carbon are within the scope of the
invention.
[0212] As used here in the term "isomer" refers to compounds having
the same molecular formula but differing in structure. Isomers
which differ only in configuration and/or conformation are referred
to as "stereoisomers." The term "isomer" is also used to refer to
an enantiomer.
[0213] The term "enantiomer" is used to describe one of a pair of
molecular isomers which are mirror images of each other and
non-superimposable. Other terms used to designate or refer to
enantiomers include "stereoisomers" (because of the different
arrangement or stereochemistry around the chiral center; although
all enantiomers are stereoisomers, not all stereoisomers are
enantiomers) or "optical isomers" (because of the optical activity
of pure enantiomers, which is the ability of different pure
enantiomers to rotate planepolarized light in different
directions). Enantiomers generally have identical physical
properties, such as melting points and boiling points, and also
have identical spectroscopic properties. Enantiomers can differ
from each other with respect to their interaction with
plane-polarized light and with respect to biological activity.
[0214] The term "racemic mixture", "racemic compound" or "racemate"
refers to a mixture of the two enantiomers of one compound. An
ideal racemic mixture is one wherein there is a 50:50 mixture of
both enantiomers of a compound such that the optical rotation of
the (+) enantiomer cancels out the optical rotation of the (-)
enantiomer.
[0215] The term "resolving" or "resolution" when used in reference
to a racemic mixture refers to the separation of a racemate into
its two enantiomorphic forms (i.e., (+) and (-); or (R) and (S)
forms). The terms can also refer to enantioselective conversion of
one isomer of a racemate to a product.
[0216] The term "enantiomeric excess" or "ee" refers to a reaction
product wherein one enantiomer is produced in excess of the other,
and is defined for a mixture of (+)- and (-)-enantiomers, with
composition given as the mole or weight or volume fraction
F.sub.(+) and F.sub.(-) (where the sum of F.sub.(+) and
F.sub.(-)=1). The enantiomeric excess is defined as *
F.sub.(+)-F.sub.(-)* and the percent enantiomeric excess by 100x*
F.sub.(+)-F.sub.(-)*. The "purity" of an enantiomer is described by
its ee or percent ee value (% ee).
[0217] Whether expressed as a "purified enantiomer" or a "pure
enantiomer" or a "resolved enantiomer" or "a compound in
enantiomeric excess", the terms are meant to indicate that the
amount of one enantiomer exceeds the amount of the other. Thus,
when referring to an enantiomer preparation, both (or either) of
the percent of the major enantiomer (e.g. by mole or by weight or
by volume) and (or) the percent enantiomeric excess of the major
enantiomer may be used to determine whether the preparation
represents a purified enantiomer preparation.
[0218] The term "enantiomeric purity" or "enantiomer purity" of an
isomer refers to a qualitative or quantitative measure of the
purified enantiomer; typically, the measurement is expressed on the
basis of ee or enantiomeric excess.
[0219] The terms "substantially purified enantiomer",
"substantially resolved enantiomer" "substantially purified
enantiomer preparation" are meant to indicate a preparation (e.g.
derived from non-optically active starting material, substrate, or
intermediate) wherein one enantiomer has been enriched over the
other, and more preferably, wherein the other enantiomer represents
less than 20%, more preferably less than 10%, and more preferably
less than 5%, and still more preferably, less than 2% of the
enantiomer or enantiomer preparation.
[0220] The terms "purified enantiomer", "resolved enantiomer" and
"purified enantiomer preparation" are meant to indicate a
preparation (e.g. derived from non-optically active starting
material, substrates or intermediates) wherein one enantiomer (for
example, the R-enantiomer) is enriched over the other, and more
preferably, wherein the other enantiomer (for example the
S-enantiomer) represents less than 30%, preferably less than 20%,
more preferably less than 10% (e.g. in this particular instance,
the R-enantiomer is substantially free of the S-enantiomer), and
more preferably less than 5% and still more preferably, less than
2% of the preparation. A purified enantiomer may be synthesized
substantially free of the other enantiomer, or a purified
enantiomer may be synthesized in a stereopreferred procedure,
followed by separation steps, or a purified enantiomer may be
derived from a racemic mixture.
[0221] The term "enantioselectivity", also called the enantiomeric
ratio indicated by the symbol "E", refers to the selective capacity
of an enzyme to generate from a racemic substrate one enantiomer
relative to the other in a product racemic mixture; in other words,
it is a measure of the ability of the enzyme to distinguish between
enantiomers. A nonselective reaction has an E of 1, while
resolutions with E's above 20 are generally considered useful for
synthesis or resolution. The enantioselectivity resides in a
difference in conversion rates between the enantiomers in question.
Reaction products are obtained that are enriched in one of the
enantiomers; conversely, remaining substrates are enriched in the
other enantiomer. For practical purposes it is generally desirable
for one of the enantiomers to be obtained in large excess. This is
achieved by terminating the conversion process at a certain degree
of conversion.
[0222] CAGE (Choline And GEranate) is an ionic liquid comprising
the cation choline (see, e.g., Structure I) and the anion geranate
or geranic acid (see, e.g., Structures II and III). Preparation of
CAGE can be, e.g., as described in International Patent Publication
WO 2015/066647; which is incorporated by reference herein in its
entirety, or as described in the examples herein.
##STR00002##
[0223] The terms "decrease", "reduced", "reduction", or "inhibit"
are all used herein to mean a decrease by a statistically
significant amount. In some embodiments, "reduce," "reduction" or
"decrease" or "inhibit" typically means a decrease by at least 10%
as compared to a reference level (e.g. the absence of a given
treatment or agent) and can include, for example, a decrease by at
least about 10%, at least about 20%, at least about 25%, at least
about 30%, at least about 35%, at least about 40%, at least about
45%, at least about 50%, at least about 55%, at least about 60%, at
least about 65%, at least about 70%, at least about 75%, at least
about 80%, at least about 85%, at least about 90%, at least about
95%, at least about 98%, at least about 99%, or more. As used
herein, "reduction" or "inhibition" does not encompass a complete
inhibition or reduction as compared to a reference level. "Complete
inhibition" is a 100% inhibition as compared to a reference level.
A decrease can be preferably down to a level accepted as within the
range of normal for an individual without a given disorder.
[0224] The terms "increased", "increase", "enhance", or "activate"
are all used herein to mean an increase by a statically significant
amount. In some embodiments, the terms "increased", "increase",
"enhance", or "activate" can mean an increase of at least 10% as
compared to a reference level, for example an increase of at least
about 20%, or at least about 30%, or at least about 40%, or at
least about 50%, or at least about 60%, or at least about 70%, or
at least about 80%, or at least about 90% or up to and including a
100% increase or any increase between 10-100% as compared to a
reference level, or at least about a 2-fold, or at least about a
3-fold, or at least about a 4-fold, or at least about a 5-fold or
at least about a 10-fold increase, or any increase between 2-fold
and 10-fold or greater as compared to a reference level. In the
context of a marker or symptom, a "increase" is a statistically
significant increase in such level.
[0225] As used herein, a "subject" means a human or animal. Usually
the animal is a vertebrate such as a primate, rodent, domestic
animal or game animal. Primates include chimpanzees, cynomologous
monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents
include mice, rats, woodchucks, ferrets, rabbits and hamsters.
Domestic and game animals include cows, horses, pigs, deer, bison,
buffalo, feline species, e.g., domestic cat, canine species, e.g.,
dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and
fish, e.g., trout, catfish and salmon. In some embodiments, the
subject is a mammal, e.g., a primate, e.g., a human. The terms,
"individual," "patient" and "subject" are used interchangeably
herein.
[0226] Preferably, the subject is a mammal. The mammal can be a
human, non-human primate, mouse, rat, dog, cat, horse, or cow, but
is not limited to these examples. Mammals other than humans can be
advantageously used as subjects that represent animal models of
conditions described herein. A subject can be male or female.
[0227] A subject can be one who has been previously diagnosed with
or identified as suffering from or having a condition in need of
treatment or one or more complications related to such a condition,
and optionally, have already undergone treatment for the condition
or the one or more complications related to the condition.
Alternatively, a subject can also be one who has not been
previously diagnosed as having the condition or one or more
complications related to the condition. For example, a subject can
be one who exhibits one or more risk factors for the condition or
one or more complications related to the condition or a subject who
does not exhibit risk factors.
[0228] A "subject in need" of treatment for a particular condition
can be a subject having that condition, diagnosed as having that
condition, or at risk of developing that condition.
[0229] As used herein, the terms "protein" and "polypeptide" are
used interchangeably herein to designate a series of amino acid
residues, connected to each other by peptide bonds between the
alpha-amino and carboxy groups of adjacent residues. The terms
"protein", and "polypeptide" refer to a polymer of amino acids,
including modified amino acids (e.g., phosphorylated, glycated,
glycosylated, etc.) and amino acid analogs, regardless of its size
or function. "Protein" and "polypeptide" are often used in
reference to relatively large polypeptides, whereas the term
"peptide" is often used in reference to small polypeptides, but
usage of these terms in the art overlaps. The terms "protein" and
"polypeptide" are used interchangeably herein when referring to a
gene product and fragments thereof. Thus, exemplary polypeptides or
proteins include gene products, naturally occurring proteins,
homologs, orthologs, paralogs, fragments and other equivalents,
variants, fragments, and analogs of the foregoing.
[0230] In the various embodiments described herein, it is further
contemplated that variants (naturally occurring or otherwise),
alleles, homologs, conservatively modified variants, and/or
conservative substitution variants of any of the particular
polypeptides described are encompassed. As to amino acid sequences,
one of skill will recognize that individual substitutions,
deletions or additions to a nucleic acid, peptide, polypeptide, or
protein sequence which alters a single amino acid or a small
percentage of amino acids in the encoded sequence is a
"conservatively modified variant" where the alteration results in
the substitution of an amino acid with a chemically similar amino
acid and retains the desired activity of the polypeptide. Such
conservatively modified variants are in addition to and do not
exclude polymorphic variants, interspecies homologs, and alleles
consistent with the disclosure.
[0231] A given amino acid can be replaced by a residue having
similar physiochemical characteristics, e.g., substituting one
aliphatic residue for another (such as Ile, Val, Leu, or Ala for
one another), or substitution of one polar residue for another
(such as between Lys and Arg; Glu and Asp; or Gln and Asn). Other
such conservative substitutions, e.g., substitutions of entire
regions having similar hydrophobicity characteristics, are well
known. Polypeptides comprising conservative amino acid
substitutions can be tested in any one of the assays described
herein to confirm that a desired activity, e.g. the activity and
specificity of a native or reference polypeptide is retained.
[0232] Amino acids can be grouped according to similarities in the
properties of their side chains (in A. L. Lehninger, in
Biochemistry, second ed., pp. 73-75, Worth Publishers, New York
(1975)): (1) non-polar: Ala (A), Val (V), Leu (L), Ile (I), Pro
(P), Phe (F), Trp (W), Met (M); (2) uncharged polar: Gly (G), Ser
(S), Thr (T), Cys (C), Tyr (Y), Asn (N), Gln (Q); (3) acidic: Asp
(D), Glu (E); (4) basic: Lys (K), Arg (R), His (H). Alternatively,
naturally occurring residues can be divided into groups based on
common side-chain properties: (1) hydrophobic: Norleucine, Met,
Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn,
Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues
that influence chain orientation: Gly, Pro; (6) aromatic: Trp, Tyr,
Phe. Non-conservative substitutions will entail exchanging a member
of one of these classes for another class. Particular conservative
substitutions include, for example; Ala into Gly or into Ser; Arg
into Lys; Asn into Gln or into His; Asp into Glu; Cys into Ser; Gln
into Asn; Glu into Asp; Gly into Ala or into Pro; His into Asn or
into Gln; Ile into Leu or into Val; Leu into Ile or into Val; Lys
into Arg, into Gln or into Glu; Met into Leu, into Tyr or into Ile;
Phe into Met, into Leu or into Tyr; Ser into Thr; Thr into Ser; Trp
into Tyr; Tyr into Trp; and/or Phe into Val, into Ile or into
Leu.
[0233] In some embodiments, the polypeptide described herein (or a
nucleic acid encoding such a polypeptide) can be a functional
fragment of one of the amino acid sequences described herein. As
used herein, a "functional fragment" is a fragment or segment of a
peptide which retains at least 50% of the wildtype reference
polypeptide's activity according to the assays described below
herein. A functional fragment can comprise conservative
substitutions of the sequences disclosed herein.
[0234] In some embodiments, the polypeptide described herein can be
a variant of a sequence described herein. In some embodiments, the
variant is a conservatively modified variant. Conservative
substitution variants can be obtained by mutations of native
nucleotide sequences, for example. A "variant," as referred to
herein, is a polypeptide substantially homologous to a native or
reference polypeptide, but which has an amino acid sequence
different from that of the native or reference polypeptide because
of one or a plurality of deletions, insertions or substitutions.
Variant polypeptide-encoding DNA sequences encompass sequences that
comprise one or more additions, deletions, or substitutions of
nucleotides when compared to a native or reference DNA sequence,
but that encode a variant protein or fragment thereof that retains
activity. A wide variety of PCR-based site-specific mutagenesis
approaches are known in the art and can be applied by the
ordinarily skilled artisan.
[0235] A variant amino acid or DNA sequence can be at least 90%, at
least 91%, at least 92%, at least 93%, at least 94%, at least 95%,
at least 96%, at least 97%, at least 98%, at least 99%, or more,
identical to a native or reference sequence. The degree of homology
(percent identity) between a native and a mutant sequence can be
determined, for example, by comparing the two sequences using
freely available computer programs commonly employed for this
purpose on the world wide web (e.g. BLASTp or BLASTn with default
settings).
[0236] In some embodiments of any of the aspects, a variant can be
a polypeptide having at least 90%, at least 95%, at least 98% or
greater sequence homology to one of the reference sequences
provided herein and retaining the wild-type activity of that
reference sequence, e.g., incretin activity. In some embodiments of
any of the aspects, a variant can be a polypeptide having at least
90%, at least 95%, at least 98% or greater sequence homology to one
of the naturally-occurring reference sequences provided herein and
retaining the wild-type activity of that reference sequence, e.g.,
incretin activity. In some embodiments of any of the aspects, a
variant can be a naturally-occurring polypeptide having at least
90%, at least 95%, at least 98% or greater sequence homology to one
of the reference sequences provided herein and retaining the
wild-type activity of that reference sequence, e.g., incretin
activity.
[0237] Alterations of the native amino acid sequence can be
accomplished by any of a number of techniques known to one of skill
in the art. Mutations can be introduced, for example, at particular
loci by synthesizing oligonucleotides containing a mutant sequence,
flanked by restriction sites enabling ligation to fragments of the
native sequence. Following ligation, the resulting reconstructed
sequence encodes an analog having the desired amino acid insertion,
substitution, or deletion. Alternatively, oligonucleotide-directed
site-specific mutagenesis procedures can be employed to provide an
altered nucleotide sequence having particular codons altered
according to the substitution, deletion, or insertion required.
Techniques for making such alterations are very well established
and include, for example, those disclosed by Walder et al. (Gene
42:133, 1986); Bauer et al. (Gene 37:73, 1985); Craik
(BioTechniques, Jan. 1985, 12-19); Smith et al. (Genetic
Engineering: Principles and Methods, Plenum Press, 1981); and U.S.
Pat. Nos. 4,518,584 and 4,737,462, which are herein incorporated by
reference in their entireties. Any cysteine residue not involved in
maintaining the proper conformation of the polypeptide also can be
substituted, generally with serine, to improve the oxidative
stability of the molecule and prevent aberrant crosslinking.
Conversely, cysteine bond(s) can be added to the polypeptide to
improve its stability or facilitate oligomerization.
[0238] As used herein, the term "antibody" refers to immunoglobulin
molecules and immunologically active portions of immunoglobulin
molecules, i.e., molecules that contain an antigen binding site
that immunospecifically binds an antigen. The term also refers to
antibodies comprised of two immunoglobulin heavy chains and two
immunoglobulin light chains as well as a variety of forms including
full length antibodies and antigen-binding portions thereof;
including, for example, an immunoglobulin molecule, a monoclonal
antibody, a chimeric antibody, a CDR-grafted antibody, a humanized
antibody, a Fab, a Fab', a F(ab')2, a Fv, a disulfide linked Fv, a
scFv, a single domain antibody (dAb), a diabody, a multispecific
antibody, a dual specific antibody, an anti-idiotypic antibody, a
bispecific antibody, a functionally active epitope-binding portion
thereof, and/or bifunctional hybrid antibodies. Each heavy chain is
composed of a variable region of said heavy chain (abbreviated here
as HCVR or VH) and a constant region of said heavy chain. The heavy
chain constant region consists of three domains CH1, CH2 and CH3.
Each light chain is composed of a variable region of said light
chain (abbreviated here as LCVR or VL) and a constant region of
said light chain. The light chain constant region consists of a CL
domain. The VH and VL regions may be further divided into
hypervariable regions referred to as complementarity-determining
regions (CDRs) and interspersed with conserved regions referred to
as framework regions (FR). Each VH and VL region thus consists of
three CDRs and four FRs which are arranged from the N terminus to
the C terminus in the following order: FR1, CDR1, FR2, CDR2, FR3,
CDR3, FR4. This structure is well known to those skilled in the
art.
[0239] As used herein, the term "antibody reagent" refers to a
polypeptide that includes at least one immunoglobulin variable
domain or immunoglobulin variable domain sequence and which
specifically binds a given antigen. An antibody reagent can
comprise an antibody or a polypeptide comprising an antigen-binding
domain of an antibody. In some embodiments, an antibody reagent can
comprise a monoclonal antibody or a polypeptide comprising an
antigen-binding domain of a monoclonal antibody. For example, an
antibody can include a heavy (H) chain variable region (abbreviated
herein as VH), and a light (L) chain variable region (abbreviated
herein as VL). In another example, an antibody includes two heavy
(H) chain variable regions and two light (L) chain variable
regions. The term "antibody reagent" encompasses antigen-binding
fragments of antibodies (e.g., single chain antibodies, Fab and
sFab fragments, F(ab')2, Fd fragments, Fv fragments, scFv, and
domain antibodies (dAb) fragments as well as complete
antibodies.
[0240] Antibodies and/or antibody reagents can include an
immunoglobulin molecule, a monoclonal antibody, a chimeric
antibody, a CDR-grafted antibody, a humanized antibody, a fully
human antibody, a Fab, a Fab', a F(ab')2, a Fv, a disulfide linked
Fv, a scFv, a single domain antibody, a diabody, a multispecific
antibody, a dual specific antibody, an anti-idiotypic antibody, a
bispecific antibody, and a functionally active epitope-binding
portion thereof.
[0241] As used herein, the term "nanobody" or single domain
antibody (sdAb) refers to an antibody comprising the small single
variable domain (WH) of antibodies obtained from camelids and
dromedaries. Antibody proteins obtained from members of the camel
and dromedary (Camelus baclrianus and Calelus dromaderius) family
including new world members such as llama species (Lama paccos,
Lama glama and Lama vicugna) have been characterized with respect
to size, structural complexity and antigenicity for human subjects.
Certain IgG antibodies from this family of mammals as found in
nature lack light chains, and are thus structurally distinct from
the typical four chain quaternary structure having two heavy and
two light chains, for antibodies from other animals. See
PCT/EP93/02214 (WO 94/04678 published 3 Mar. 1994; which is
incorporated by reference herein in its entirety).
[0242] A region of the camelid antibody which is the small single
variable domain identified as VHH can be obtained by genetic
engineering to yield a small protein having high affinity for a
target, resulting in a low molecular weight antibody-derived
protein known as a "camelid nanobody". See U.S. Pat. No. 5,759,808
issued Jun. 2, 1998; see also Stijlemans, B. et al., 2004 J Biol
Chem 279: 1256-1261; Dumoulin, M. et al., 2003 Nature 424: 783-788;
Pleschberger, M. et al. 2003 Bioconjugate Chem 14: 440-448;
Cortez-Retamozo, V. et al. 2002 Int J Cancer 89: 456-62; and
Lauwereys, M. et al. 1998 EMBO J. 17: 3512-3520; each of which is
incorporated by reference herein in its entirety. Engineered
libraries of camelid antibodies and antibody fragments are
commercially available, for example, from Ablynx, Ghent, Belgium.
As with other antibodies of non-human origin, an amino acid
sequence of a camelid antibody can be altered recombinantly to
obtain a sequence that more closely resembles a human sequence,
i.e., the nanobody can be "humanized". Thus the natural low
antigenicity of camelid antibodies to humans can be further
reduced.
[0243] The camelid nanobody has a molecular weight approximately
one-tenth that of a human IgG molecule and the protein has a
physical diameter of only a few nanometers. One consequence of the
small size is the ability of camelid nanobodies to bind to
antigenic sites that are functionally invisible to larger antibody
proteins, i.e., camelid nanobodies are useful as reagents detect
antigens that are otherwise cryptic using classical immunological
techniques, and as possible therapeutic agents. Thus yet another
consequence of small size is that a camelid nanobody can inhibit as
a result of binding to a specific site in a groove or narrow cleft
of a target protein, and hence can serve in a capacity that more
closely resembles the function of a classical low molecular weight
drug than that of a classical antibody. The low molecular weight
and compact size further result in camelid nanobodies being
extremely thermostable, stable to extreme pH and to proteolytic
digestion, and poorly antigenic. See U.S. patent application
20040161738 published Aug. 19, 2004; which is incorporated by
reference herein in its entirety. These features combined with the
low antigenicity to humans indicate great therapeutic
potential.
[0244] As used herein, the term "nucleic acid" or "nucleic acid
sequence" refers to any molecule, preferably a polymeric molecule,
incorporating units of ribonucleic acid, deoxyribonucleic acid or
an analog thereof. The nucleic acid can be either single-stranded
or double-stranded. A single-stranded nucleic acid can be one
nucleic acid strand of a denatured double-stranded DNA.
Alternatively, it can be a single-stranded nucleic acid not derived
from any double-stranded DNA. In one aspect, the nucleic acid can
be DNA. In another aspect, the nucleic acid can be RNA. Suitable
DNA can include, e.g., cDNA. Suitable RNA can include, e.g.,
mRNA.
[0245] As used herein, "inhibitory nucleic acid" refers to a
nucleic acid molecule which can inhibit the expression of a target,
e.g., double-stranded RNAs (dsRNAs), inhibitory RNAs (iRNAs), and
the like.
[0246] Double-stranded RNA molecules (dsRNA) have been shown to
block gene expression in a highly conserved regulatory mechanism
known as RNA interference (RNAi). The inhibitory nucleic acids
described herein can include an RNA strand (the antisense strand)
having a region which is 30 nucleotides or less in length, i.e.,
15-30 nucleotides in length, generally 19-24 nucleotides in length,
which region is substantially complementary to at least part the
targeted mRNA transcript. The use of these iRNAs enables the
targeted degradation of mRNA transcripts, resulting in decreased
expression and/or activity of the target.
[0247] As used herein, the term "iRNA" refers to an agent that
contains RNA (or modified nucleic acids as described below herein)
and which mediates the targeted cleavage of an RNA transcript via
an RNA-induced silencing complex (RISC) pathway. In some
embodiments of any of the aspects, an iRNA as described herein
effects inhibition of the expression and/or activity of a target.
In some embodiments of any of the aspects, contacting a cell with
the inhibitor (e.g. an iRNA) results in a decrease in the target
mRNA level in a cell by at least about 5%, about 10%, about 20%,
about 30%, about 40%, about 50%, about 60%, about 70%, about 80%,
about 90%, about 95%, about 99%, up to and including 100% of the
target mRNA level found in the cell without the presence of the
iRNA. In some embodiments of any of the aspects, administering an
inhibitor (e.g. an iRNA) to a subject results in a decrease in the
target mRNA level in the subject by at least about 5%, about 10%,
about 20%, about 30%, about 40%, about 50%, about 60%, about 70%,
about 80%, about 90%, about 95%, about 99%, up to and including
100% of the target mRNA level found in the subject without the
presence of the iRNA.
[0248] In some embodiments of any of the aspects, the iRNA can be a
dsRNA. A dsRNA includes two RNA strands that are sufficiently
complementary to hybridize to form a duplex structure under
conditions in which the dsRNA will be used. One strand of a dsRNA
(the antisense strand) includes a region of complementarity that is
substantially complementary, and generally fully complementary, to
a target sequence. The target sequence can be derived from the
sequence of an mRNA formed during the expression of the target,
e.g., it can span one or more intron boundaries. The other strand
(the sense strand) includes a region that is complementary to the
antisense strand, such that the two strands hybridize and form a
duplex structure when combined under suitable conditions.
Generally, the duplex structure is between 15 and 30 base pairs in
length inclusive, more generally between 18 and 25 base pairs in
length inclusive, yet more generally between 19 and 24 base pairs
in length inclusive, and most generally between 19 and 21 base
pairs in length, inclusive. Similarly, the region of
complementarity to the target sequence is between 15 and 30 base
pairs in length inclusive, more generally between 18 and 25 base
pairs in length inclusive, yet more generally between 19 and 24
base pairs in length inclusive, and most generally between 19 and
21 base pairs in length nucleotides in length, inclusive. In some
embodiments of any of the aspects, the dsRNA is between 15 and 20
nucleotides in length, inclusive, and in other embodiments, the
dsRNA is between 25 and 30 nucleotides in length, inclusive. As the
ordinarily skilled person will recognize, the targeted region of an
RNA targeted for cleavage will most often be part of a larger RNA
molecule, often an mRNA molecule. Where relevant, a "part" of an
mRNA target is a contiguous sequence of an mRNA target of
sufficient length to be a substrate for RNAi-directed cleavage
(i.e., cleavage through a RISC pathway). dsRNAs having duplexes as
short as 9 base pairs can, under some circumstances, mediate
RNAi-directed RNA cleavage. Most often a target will be at least 15
nucleotides in length, preferably 15-30 nucleotides in length.
[0249] Exemplary embodiments of types of inhibitory nucleic acids
can include, e.g,. siRNA, shRNA, miRNA, and/or amiRNA, which are
well known in the art.
[0250] In some embodiments of any of the aspects, the RNA of an
iRNA, e.g., a dsRNA, is chemically modified to enhance stability or
other beneficial characteristics. The nucleic acids described
herein may be synthesized and/or modified by methods well
established in the art, such as those described in "Current
protocols in nucleic acid chemistry," Beaucage, S. L. et al.
(Edrs.), John Wiley & Sons, Inc., New York, N.Y., USA, which is
hereby incorporated herein by reference. Modifications include, for
example, (a) end modifications, e.g., 5' end modifications
(phosphorylation, conjugation, inverted linkages, etc.) 3' end
modifications (conjugation, DNA nucleotides, inverted linkages,
etc.), (b) base modifications, e.g., replacement with stabilizing
bases, destabilizing bases, or bases that base pair with an
expanded repertoire of partners, removal of bases (abasic
nucleotides), or conjugated bases, (c) sugar modifications (e.g.,
at the 2' position or 4' position) or replacement of the sugar, as
well as (d) backbone modifications, including modification or
replacement of the phosphodiester linkages. Specific examples of
RNA compounds useful in the embodiments described herein include,
but are not limited to RNAs containing modified backbones or no
natural internucleoside linkages. RNAs having modified backbones
include, among others, those that do not have a phosphorus atom in
the backbone. For the purposes of this specification, and as
sometimes referenced in the art, modified RNAs that do not have a
phosphorus atom in their internucleoside backbone can also be
considered to be oligonucleosides. In some embodiments of any of
the aspects, the modified RNA will have a phosphorus atom in its
internucleoside backbone.
[0251] Modified RNA backbones can include, for example,
phosphorothioates, chiral phosphorothioates, phosphorodithioates,
phosphotriesters, aminoalkylphosphotriesters, methyl and other
alkyl phosphonates including 3'-alkylene phosphonates and chiral
phosphonates, phosphinates, phosphoramidates including 3'-amino
phosphoramidate and aminoalkylphosphoramidates,
thionophosphoramidates, thionoalkylphosphonates,
thionoalkylphosphotriesters, and boranophosphates having normal
3'-5' linkages, 2'-5' linked analogs of these, and those) having
inverted polarity wherein the adjacent pairs of nucleoside units
are linked 3'-5' to 5'-3' or 2'-5' to 5'-2'. Various salts, mixed
salts and free acid forms are also included. Modified RNA backbones
that do not include a phosphorus atom therein have backbones that
are formed by short chain alkyl or cycloalkyl internucleoside
linkages, mixed heteroatoms and alkyl or cycloalkyl internucleoside
linkages, or one or more short chain heteroatomic or heterocyclic
internucleoside linkages. These include those having morpholino
linkages (formed in part from the sugar portion of a nucleoside);
siloxane backbones; sulfide, sulfoxide and sulfone backbones;
formacetyl and thioformacetyl backbones; methylene formacetyl and
thioformacetyl backbones; alkene containing backbones; sulfamate
backbones; methyleneimino and methylenehydrazino backbones;
sulfonate and sulfonamide backbones; amide backbones; others having
mixed N, O, S and CH2 component parts, and oligonucleosides with
heteroatom backbones, and in particular --CH2-NH--CH2-,
--CH2-N(CH3)-O--CH2-[known as a methylene (methylimino) or MMI
backbone], --CH2-O--N(CH3)-CH2-, --CH2-N(CH3)-N(CH3)-CH2- and
--N(CH3)-CH2-CH2-[wherein the native phosphodiester backbone is
represented as --O--P--O--CH2-].
[0252] In other RNA mimetics suitable or contemplated for use in
iRNAs, both the sugar and the internucleoside linkage, i.e., the
backbone, of the nucleotide units are replaced with novel groups.
The base units are maintained for hybridization with an appropriate
nucleic acid target compound. One such oligomeric compound, an RNA
mimetic that has been shown to have excellent hybridization
properties, is referred to as a peptide nucleic acid (PNA). In PNA
compounds, the sugar backbone of an RNA is replaced with an amide
containing backbone, in particular an aminoethylglycine backbone.
The nucleobases are retained and are bound directly or indirectly
to aza nitrogen atoms of the amide portion of the backbone.
[0253] The RNA of an iRNA can also be modified to include one or
more locked nucleic acids (LNA). A locked nucleic acid is a
nucleotide having a modified ribose moiety in which the ribose
moiety comprises an extra bridge connecting the 2' and 4' carbons.
This structure effectively "locks" the ribose in the 3'-endo
structural conformation. The addition of locked nucleic acids to
siRNAs has been shown to increase siRNA stability in serum, and to
reduce off-target effects (Elmen, J. et al., (2005) Nucleic Acids
Research 33(1):439-447; Mook, O R. et al., (2007) Mol Canc Ther
6(3):833-843; Grunweller, A. et al., (2003) Nucleic Acids Research
31(12):3185-3193).
[0254] Modified RNAs can also contain one or more substituted sugar
moieties. The iRNAs, e.g., dsRNAs, described herein can include one
of the following at the 2' position: OH; F; O-, S-, or N-alkyl; O-,
S-, or N-alkenyl; O-, S- or N-alkynyl; or O-alkyl-O-alkyl, wherein
the alkyl, alkenyl and alkynyl may be substituted or unsubstituted
C1 to C10 alkyl or C2 to C10 alkenyl and alkynyl. Exemplary
suitable modifications include O[(CH2)nO] mCH3, O(CH2).nOCH3,
O(CH2)nNH2, O(CH2) nCH3, O(CH2)nONH2, and
O(CH.sub.2)nON[(CH2)nCH3)]2, where n and m are from 1 to about 10.
In some embodiments of any of the aspects, dsRNAs include one of
the following at the 2' position: C1 to C10 lower alkyl,
substituted lower alkyl, alkaryl, aralkyl, O-alkaryl or O-aralkyl,
SH, SCH3, OCN, Cl, Br, CN, CF.sub.3, OCF3, SOCH3, SO2CH3, ONO2,
NO2, N3, NH2, heterocycloalkyl, heterocycloalkaryl,
aminoalkylamino, polyalkylamino, substituted silyl, an RNA cleaving
group, a reporter group, an intercalator, a group for improving the
pharmacokinetic properties of an iRNA, or a group for improving the
pharmacodynamic properties of an iRNA, and other substituents
having similar properties. In some embodiments of any of the
aspects, the modification includes a 2' methoxyethoxy
(2'-O--CH2CH2OCH3, also known as 2'-O-(2-methoxyethyl) or 2'-MOE)
(Martin et al., Helv. Chim. Acta, 1995, 78:486-504) i.e., an
alkoxy-alkoxy group. Another exemplary modification is
2'-dimethylaminooxyethoxy, i.e., a O(CH2)2ON(CH3)2 group, also
known as 2'-DMAOE, as described in examples herein below, and
2'-dimethylaminoethoxyethoxy (also known in the art as
2'-O-dimethylaminoethoxyethyl or 2'-DMAEOE), i.e.,
2'-O--CH2-O--CH2-N(CH2)2, also described in examples herein
below.
[0255] Other modifications include 2'-methoxy (2'-OCH3),
2'-aminopropoxy (2'-OCH2CH2CH2NH2) and 2'-fluoro (2'-F). Similar
modifications can also be made at other positions on the RNA of an
iRNA, particularly the 3' position of the sugar on the 3' terminal
nucleotide or in 2'-5' linked dsRNAs and the 5' position of 5'
terminal nucleotide. iRNAs may also have sugar mimetics such as
cyclobutyl moieties in place of the pentofuranosyl sugar.
[0256] An inhibitory nucleic acid can also include nucleobase
(often referred to in the art simply as "base") modifications or
substitutions. As used herein, "unmodified" or "natural"
nucleobases include the purine bases adenine (A) and guanine (G),
and the pyrimidine bases thymine (T), cytosine (C) and uracil (U).
Modified nucleobases include other synthetic and natural
nucleobases such as 5-methylcytosine (5-me-C), 5-hydroxymethyl
cytosine, xanthine, hypoxanthine, 2-aminoadenine, 6-methyl and
other alkyl derivatives of adenine and guanine, 2-propyl and other
alkyl derivatives of adenine and guanine, 2-thiouracil,
2-thiothymine and 2-thiocytosine, 5-halouracil and cytosine,
5-propynyl uracil and cytosine, 6-azo uracil, cytosine and thymine,
5-uracil (pseudouracil), 4-thiouracil, 8-halo, 8-amino, 8-thiol,
8-thioalkyl, 8-hydroxyl anal other 8-substituted adenines and
guanines, 5-halo, particularly 5-bromo, 5-trifluoromethyl and other
5-substituted uracils and cytosines, 7-methylguanine and
7-methyladenine, 8-azaguanine and 8-azaadenine, 7-deazaguanine and
7-daazaadenine and 3-deazaguanine and 3-deazaadenine. Certain of
these nucleobases are particularly useful for increasing the
binding affinity of the inhibitory nucleic acids featured in the
invention. These include 5-substituted pyrimidines,
6-azapyrimidines and N-2, N-6 and 0-6 substituted purines,
including 2-aminopropyladenine, 5-propynyluracil and
5-propynylcytosine. 5-methylcytosine substitutions have been shown
to increase nucleic acid duplex stability by 0.6-1.2.degree. C.
(Sanghvi, Y. S., Crooke, S. T. and Lebleu, B., Eds., dsRNA Research
and Applications, CRC Press, Boca Raton, 1993, pp. 276-278) and are
exemplary base substitutions, even more particularly when combined
with 2'-O-methoxyethyl sugar modifications.
[0257] The preparation of the modified nucleic acids, backbones,
and nucleobases described above are well known in the art.
[0258] Another modification of an inhibitory nucleic acid featured
in the invention involves chemically linking to the inhibitory
nucleic acid to one or more ligands, moieties or conjugates that
enhance the activity, cellular distribution, pharmacokinetic
properties, or cellular uptake of the iRNA. Such moieties include
but are not limited to lipid moieties such as a cholesterol moiety
(Letsinger et al., Proc. Natl. Acid. Sci. USA, 1989, 86:
6553-6556), cholic acid (Manoharan et al., Biorg. Med. Chem. Let.,
1994, 4:1053-1060), a thioether, e.g., beryl-S-tritylthiol
(Manoharan et al., Ann. N.Y. Acad. Sci., 1992, 660:306-309;
Manoharan et al., Biorg. Med. Chem. Let., 1993, 3:2765-2770), a
thiocholesterol (Oberhauser et al., Nucl. Acids Res., 1992,
20:533-538), an aliphatic chain, e.g., dodecandiol or undecyl
residues (Saison-Behmoaras et al., EMBO J, 1991, 10:1111-1118;
Kabanov et al., FEBS Lett., 1990, 259:327-330; Svinarchuk et al.,
Biochimie, 1993, 75:49-54), a phospholipid, e.g.,
di-hexadecyl-rac-glycerol or triethyl-ammonium
1,2-di-O-hexadecyl-rac-glycero-3-phosphonate (Manoharan et al.,
Tetrahedron Lett., 1995, 36:3651-3654; Shea et al., Nucl. Acids
Res., 1990, 18:3777-3783), a polyamine or a polyethylene glycol
chain (Manoharan et al., Nucleosides & Nucleotides, 1995,
14:969-973), or adamantane acetic acid (Manoharan et al.,
Tetrahedron Lett., 1995, 36:3651-3654), a palmityl moiety (Mishra
et al., Biochim. Biophys. Acta, 1995, 1264:229-237), or an
octadecylamine or hexylamino-carbonyloxycholesterol moiety (Crooke
et al., J. Pharmacol. Exp. Ther., 1996, 277:923-937).
[0259] The term "vector", as used herein, refers to a nucleic acid
construct designed for delivery to a host cell or for transfer
between different host cells. As used herein, a vector can be viral
or non-viral. The term "vector" encompasses any genetic element
that is capable of replication when associated with the proper
control elements and that can transfer gene sequences to cells. A
vector can include, but is not limited to, a cloning vector, an
expression vector, a recombinant vector, a plasmid, phage,
transposon, cosmid, chromosome, virus, virion, etc.
[0260] As used herein, the term "expression vector" refers to a
vector that directs expression of an RNA or polypeptide from
sequences linked to transcriptional regulatory sequences on the
vector. The sequences expressed will often, but not necessarily, be
heterologous to the cell. An expression vector may comprise
additional elements, for example, the expression vector may have
two replication systems, thus allowing it to be maintained in two
organisms, for example in human cells for expression and in a
prokaryotic host for cloning and amplification. The term
"expression" refers to the cellular processes involved in producing
RNA and proteins and as appropriate, secreting proteins, including
where applicable, but not limited to, for example, transcription,
transcript processing, translation and protein folding,
modification and processing. "Expression products" include RNA
transcribed from a gene, and polypeptides obtained by translation
of mRNA transcribed from a gene. The term "gene" means the nucleic
acid sequence which is transcribed (DNA) to RNA in vitro or in vivo
when operably linked to appropriate regulatory sequences. The gene
may or may not include regions preceding and following the coding
region, e.g. 5' untranslated (5'UTR) or "leader" sequences and 3'
UTR or "trailer" sequences, as well as intervening sequences
(introns) between individual coding segments (exons).
[0261] As used herein, the term "viral vector" refers to a nucleic
acid vector construct that includes at least one element of viral
origin and has the capacity to be packaged into a viral vector
particle. The viral vector can contain the nucleic acid encoding a
polypeptide as described herein in place of non-essential viral
genes. The vector and/or particle may be utilized for the purpose
of transferring any nucleic acids into cells either in vitro or in
vivo. Numerous forms of viral vectors are known in the art.
[0262] By "recombinant vector" is meant a vector that includes a
heterologous nucleic acid sequence, or "transgene" that is capable
of expression in vivo. It should be understood that the vectors
described herein can, in some embodiments, be combined with other
suitable compositions and therapies. In some embodiments, the
vector is episomal. The use of a suitable episomal vector provides
a means of maintaining the nucleotide of interest in the subject in
high copy number extra chromosomal DNA thereby eliminating
potential effects of chromosomal integration.
[0263] As used herein, the terms "treat," "treatment," "treating,"
or "amelioration" refer to therapeutic treatments, wherein the
object is to reverse, alleviate, ameliorate, inhibit, slow down or
stop the progression or severity of a condition associated with a
disease or disorder, e.g. a condition or disease described herein.
The term "treating" includes reducing or alleviating at least one
adverse effect or symptom of a condition, disease or disorder.
Treatment is generally "effective" if one or more symptoms or
clinical markers are reduced. Alternatively, treatment is
"effective" if the progression of a disease is reduced or halted.
That is, "treatment" includes not just the improvement of symptoms
or markers, but also a cessation of, or at least slowing of,
progress or worsening of symptoms compared to what would be
expected in the absence of treatment. Beneficial or desired
clinical results include, but are not limited to, alleviation of
one or more symptom(s), diminishment of extent of disease,
stabilized (i.e., not worsening) state of disease, delay or slowing
of disease progression, amelioration or palliation of the disease
state, remission (whether partial or total), and/or decreased
mortality, whether detectable or undetectable. The term "treatment"
of a disease also includes providing relief from the symptoms or
side-effects of the disease (including palliative treatment).
[0264] As used herein, the term "pharmaceutical composition" refers
to the active agent in combination with a pharmaceutically
acceptable carrier e.g. a carrier commonly used in the
pharmaceutical industry. The phrase "pharmaceutically acceptable"
is employed herein to refer to those compounds, materials,
compositions, and/or dosage forms which are, within the scope of
sound medical judgment, suitable for use in contact with the
tissues of human beings and animals without excessive toxicity,
irritation, allergic response, or other problem or complication,
commensurate with a reasonable benefit/risk ratio. In some
embodiments of any of the aspects, a pharmaceutically acceptable
carrier can be a carrier other than water. In some embodiments of
any of the aspects, a pharmaceutically acceptable carrier can be a
cream, emulsion, gel, liposome, nanoparticle, and/or ointment. In
some embodiments of any of the aspects, a pharmaceutically
acceptable carrier can be an artificial or engineered carrier,
e.g., a carrier that the active ingredient would not be found to
occur in in nature.
[0265] As used herein, the term "administering," refers to the
placement of a compound as disclosed herein into a subject by a
method or route which results in at least partial delivery of the
agent at a desired site. Pharmaceutical compositions comprising the
compounds disclosed herein can be administered by any appropriate
route which results in an effective treatment in the subject.
[0266] As used herein, "contacting" refers to any suitable means
for delivering, or exposing, an agent to at least one cell.
Exemplary delivery methods include, but are not limited to, direct
delivery to cell culture medium, perfusion, injection, or other
delivery method well known to one skilled in the art. In some
embodiments, contacting comprises physical human activity, e.g., an
injection; an act of dispensing, mixing, and/or decanting; and/or
manipulation of a delivery device or machine.
[0267] The term "effective amount" means an amount of a composition
sufficient to provide at least some amelioration of the symptoms
associated with the condition. In one embodiment, the "effective
amount" means an amount of a composition would decrease the markers
or symptoms of the condition in a subject having the condition.
[0268] The term "statistically significant" or "significantly"
refers to statistical significance and generally means a two
standard deviation (2SD) or greater difference.
[0269] Other than in the operating examples, or where otherwise
indicated, all numbers expressing quantities of ingredients or
reaction conditions used herein should be understood as modified in
all instances by the term "about." The term "about" when used in
connection with percentages can mean.+-.1%.
[0270] As used herein, the term "comprising" or "comprises" is used
in reference to methods and compositions, and respective
component(s) thereof, that are essential to the invention, yet open
to the inclusion of unspecified elements, whether essential or not.
As used herein, the term "comprising" means that other elements can
also be present in addition to the defined elements presented. The
use of "comprising" indicates inclusion rather than limitation.
[0271] The term "consisting of" refers to compositions, methods,
and respective components thereof as described herein, which are
exclusive of any element not recited in that description of the
embodiment.
[0272] As used herein the term "consisting essentially of" refers
to those elements required for a given embodiment. The term permits
the presence of additional elements that do not materially affect
the basic and novel or functional characteristic(s) of that
embodiment of the invention.
[0273] As used herein, the term "specific binding" refers to a
chemical interaction between two molecules, compounds, cells and/or
particles wherein the first entity binds to the second, target
entity with greater specificity and affinity than it binds to a
third entity which is a non-target. In some embodiments, specific
binding can refer to an affinity of the first entity for the second
target entity which is at least 10 times, at least 50 times, at
least 100 times, at least 500 times, at least 1000 times or greater
than the affinity for the third nontarget entity. A reagent
specific for a given target is one that exhibits specific binding
for that target under the conditions of the assay being
utilized.
[0274] The singular terms "a," "an," and "the" include plural
referents unless context clearly indicates otherwise. Similarly,
the word "or" is intended to include "and" unless the context
clearly indicates otherwise. Although methods and materials similar
or equivalent to those described herein can be used in the practice
or testing of this disclosure, suitable methods and materials are
described below. The abbreviation, "e.g." is derived from the Latin
exempli gratia, and is used herein to indicate a non-limiting
example. Thus, the abbreviation "e.g." is synonymous with the term
"for example."
[0275] Groupings of alternative elements or embodiments of the
invention disclosed herein are not to be construed as limitations.
Each group member can be referred to and claimed individually or in
any combination with other members of the group or other elements
found herein. One or more members of a group can be included in, or
deleted from, a group for reasons of convenience and/or
patentability. When any such inclusion or deletion occurs, the
specification is herein deemed to contain the group as modified
thus fulfilling the written description of all Markush groups used
in the appended claims.
[0276] Unless otherwise defined herein, scientific and technical
terms used in connection with the present application shall have
the meanings that are commonly understood by those of ordinary
skill in the art to which this disclosure belongs. It should be
understood that this invention is not limited to the particular
methodology, protocols, and reagents, etc., described herein and as
such can vary. The terminology used herein is for the purpose of
describing particular embodiments only, and is not intended to
limit the scope of the present invention, which is defined solely
by the claims. Definitions of common terms in immunology and
molecular biology can be found in The Merck Manual of Diagnosis and
Therapy, 19th Edition, published by Merck Sharp & Dohme Corp.,
2011 (ISBN 978-0-911910-19-3); Robert S. Porter et al. (eds.), The
Encyclopedia of Molecular Cell Biology and Molecular Medicine,
published by Blackwell Science Ltd., 1999-2012 (ISBN
9783527600908); and Robert A. Meyers (ed.), Molecular Biology and
Biotechnology: a Comprehensive Desk Reference, published by VCH
Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Werner
Luttmann, published by Elsevier, 2006; Janeway's Immunobiology,
Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), Taylor &
Francis Limited, 2014 (ISBN 0815345305, 9780815345305); Lewin's
Genes XI, published by Jones & Bartlett Publishers, 2014
(ISBN-1449659055); Michael Richard Green and Joseph Sambrook,
Molecular Cloning: A Laboratory Manual, 4.sup.th ed., Cold Spring
Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN
1936113414); Davis et al., Basic Methods in Molecular Biology,
Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN
044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch
(ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in
Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley
and Sons, 2014 (ISBN 047150338X, 9780471503385), Current Protocols
in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and
Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John
E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach,
Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN
0471142735, 9780471142737), the contents of which are all
incorporated by reference herein in their entireties.
[0277] One of skill in the art can readily identify a
chemotherapeutic agent of use (e.g. see Physicians' Cancer
Chemotherapy Drug Manual 2014, Edward Chu, Vincent T. DeVita Jr.,
Jones & Bartlett Learning; Principles of Cancer Therapy,
Chapter 85 in Harrison's Principles of Internal Medicine, 18th
edition; Therapeutic Targeting of Cancer Cells: Era of Molecularly
Targeted Agents and Cancer Pharmacology, Chs. 28-29 in Abeloff's
Clinical Oncology, 2013 Elsevier; and Fischer D S (ed): The Cancer
Chemotherapy Handbook, 4th ed. St. Louis, Mosby-Year Book,
2003).
[0278] Other terms are defined herein within the description of the
various aspects of the invention.
[0279] All patents and other publications; including literature
references, issued patents, published patent applications, and
co-pending patent applications; cited throughout this application
are expressly incorporated herein by reference for the purpose of
describing and disclosing, for example, the methodologies described
in such publications that might be used in connection with the
technology described herein. These publications are provided solely
for their disclosure prior to the filing date of the present
application. Nothing in this regard should be construed as an
admission that the inventors are not entitled to antedate such
disclosure by virtue of prior invention or for any other reason.
All statements as to the date or representation as to the contents
of these documents is based on the information available to the
applicants and does not constitute any admission as to the
correctness of the dates or contents of these documents.
[0280] The description of embodiments of the disclosure is not
intended to be exhaustive or to limit the disclosure to the precise
form disclosed. While specific embodiments of, and examples for,
the disclosure are described herein for illustrative purposes,
various equivalent modifications are possible within the scope of
the disclosure, as those skilled in the relevant art will
recognize. For example, while method steps or functions are
presented in a given order, alternative embodiments may perform
functions in a different order, or functions may be performed
substantially concurrently. The teachings of the disclosure
provided herein can be applied to other procedures or methods as
appropriate. The various embodiments described herein can be
combined to provide further embodiments. Aspects of the disclosure
can be modified, if necessary, to employ the compositions,
functions and concepts of the above references and application to
provide yet further embodiments of the disclosure. Moreover, due to
biological functional equivalency considerations, some changes can
be made in protein structure without affecting the biological or
chemical action in kind or amount. These and other changes can be
made to the disclosure in light of the detailed description. All
such modifications are intended to be included within the scope of
the appended claims.
[0281] Specific elements of any of the foregoing embodiments can be
combined or substituted for elements in other embodiments.
Furthermore, while advantages associated with certain embodiments
of the disclosure have been described in the context of these
embodiments, other embodiments may also exhibit such advantages,
and not all embodiments need necessarily exhibit such advantages to
fall within the scope of the disclosure.
[0282] The technology described herein is further illustrated by
the following examples which in no way should be construed as being
further limiting.
[0283] Some embodiments of the technology described herein can be
defined according to any of the following numbered paragraphs:
[0284] 1. A method of administering at least one active compound,
the method comprising administering the active compound in
combination with at least one ionic liquid comprising: [0285] a
hydrophobic anion comprising a carboxylic acid having a pKa of at
least 4.0 and a Log P of at least 1.0; and [0286] a cation
comprising a quaternary ammonium. [0287] 2. A method of reducing
weight/weight gain or treating obesity, diabetes, ulcers, cancer,
or fibrosis in a subject in need thereof, the method comprising
administering a composition comprising at least one ionic liquid
comprising: [0288] a hydrophobic anion comprising a carboxylic acid
having a pKa of at least 4.0 and a Log P of at least 1.0; and
[0289] a cation comprising a quaternary ammonium to the subject.
[0290] 3. The method of paragraph 2, wherein the composition does
not comprise a therapeutic agent other than the at least one ionic
liquid. [0291] 4. The method of paragraph 2, wherein the
composition further comprises an active compound other than the at
least one ionic liquid. [0292] 5. The method of any of the
preceding paragraphs, wherein the anion has a pKa of at least 4.5.
[0293] 6. The method of any of the preceding paragraphs, wherein
the anion has a pKa of at least 5.0. [0294] 7. The method of any of
the preceding paragraphs, wherein the anion has a Log P of at least
2.0. [0295] 8. The method of any of the preceding paragraphs,
wherein the anion has a Log P of at least 2.5. [0296] 9. The method
of any of the preceding paragraphs, wherein the anion has a Log P
of at least 2.75. [0297] 10. The method of any of the preceding
paragraphs, wherein the anion comprises a carbon chain of at least
8 carbons. [0298] 11. The method of any of the preceding
paragraphs, wherein the anion is an alkene. [0299] 12. The method
of any of the preceding paragraphs, wherein the anion is geranic
acid, octanoic acid, or citronellic acid. [0300] 13. The method of
any of the preceding paragraphs, wherein the cation has a molar
mass equal to or greater than choline. [0301] 14. The method of any
of the preceding paragraphs, wherein the quarternary ammonium has
the structure of NR.sub.4.sup.+ and at least one R group comprises
a hydroxy group. [0302] 15. The method of any of the preceding
paragraphs, wherein the quarternary ammonium has the structure of
NR.sub.4.sup.+ and only one R group comprises a hydroxy group.
[0303] 16. The method of any of the preceding paragraphs, wherein
the cation is C1, C6, or C7. [0304] 17. The method of any of the
preceding paragraphs, wherein the cation is selected from choline,
C1, C6, and C7 and the anion is citronellic acid. [0305] 18. The
method of any of the preceding paragraphs, wherein the cation is C1
and the anion is citronellic acid. [0306] 19. The method of any of
the preceding paragraphs, wherein the cation is selected from C1,
C6, and C7 and the anion is geranic acid. [0307] 20. The method of
any of the preceding paragraphs, wherein the ionic liquid is
choline: citronellic acid, C1: geranic acid, or C1: citronellic
acid. [0308] 21. The method of any of the preceding paragraphs,
wherein the ionic liquid is not CAGE. [0309] 22. The method of any
of the preceding paragraphs, wherein the ionic liquid has less than
20 cross peaks as measured by Nuclear Overhauser Effect
SpectroscopY (NOESY). [0310] 23. The method of any of the preceding
paragraphs, wherein the ionic liquid has less than 10 cross peaks
as measured by Nuclear Overhauser Effect SpectroscopY (NOESY).
[0311] 24. The method of any of the preceding paragraphs, wherein
the ionic liquid has less than 5 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY). [0312] 25. The method of
any of the preceding paragraphs, wherein the administration is
transdermal. [0313] 26. The method of any of the preceding
paragraphs, wherein the administration is transdermal, to a mucus
membrane, oral, subcutaneous, intradermal, parenteral,
intratumoral, or intravenous. [0314] 27. The method of paragraph
26, wherein the mucus membrane is nasal, oral, or vaginal. [0315]
28. The method of any of the preceding paragraphs, wherein the
ionic liquid is at a concentration of at least 0.1% w/v. [0316] 29.
The method of any of the preceding paragraphs, wherein the ionic
liquid is at a concentration of from about 10 to about 70% w/v.
[0317] 30. The method of any of the preceding paragraphs, wherein
the ionic liquid is at a concentration of from about 30 to about
50% w/v. [0318] 31. The method of any of the preceding paragraphs,
wherein the ionic liquid is at a concentration of from about 30 to
about 40% w/v. [0319] 32. The method of any of the preceding
paragraphs, wherein the ionic liquid comprises a ratio of cation to
anion of from about 2:1 to about 1:10. [0320] 33. The method of any
of the preceding paragraphs, wherein the ionic liquid comprises a
ratio of cation to anion of from about 1:1 to about 1:4. [0321] 34.
The method of any of the preceding paragraphs, wherein the ionic
liquid comprises a ratio of cation to anion of about 1:2. [0322]
35. The method of any of the preceding paragraphs, wherein the
ionic liquid has a cation:anion ratio of less than 1:1. [0323] 36.
The method of any of the preceding paragraphs, wherein the active
compound is hydrophobic. [0324] 37. The method of any of the
preceding paragraphs, wherein the active compound is hydrophilic.
[0325] 38. The method of any of the preceding paragraphs, wherein
the active compound comprises a polypeptide. [0326] 39. The method
of any of the preceding paragraphs, wherein the active compound has
a molecular weight of greater than 450. [0327] 40. The method of
any of the preceding paragraphs, wherein the active compound has a
molecular weight of greater than 500. [0328] 41. The method of any
of the preceding paragraphs, wherein the active compound comprises
an antibody or antibody reagent. [0329] 42. The method of any of
the preceding paragraphs, wherein the active compound comprises
insulin, acarbose, ruxolitinib, or a GLP-1 polypeptide or mimetic
or analog thereof [0330] 43. The method of any of the preceding
paragraphs, wherein the combination and/or composition is
administered once. [0331] 44. The method of any of the preceding
paragraphs, wherein the combination and/or composition is
administered in multiple doses. [0332] 45. The method of any of the
preceding paragraphs, wherein the active compound and/or
composition is provided at a dosage of 1-20 mg/kg. [0333] 46. The
method of any of the preceding paragraphs, wherein the active
compound and the ionic liquid are further in combination with at
least one non-ionic surfactant. [0334] 47. The method of any of the
preceding paragraphs, wherein the combination and/or composition
further comprises a further pharmaceutically acceptable carrier.
[0335] 48. The method of any of the preceding paragraphs, wherein
the administration is oral and the combination and/or composition
is provided in a degradable capsule. [0336] 49. The method of any
of the preceding paragraphs, wherein the combination is an
admixture. [0337] 50. The method of any of the preceding
paragraphs, wherein the combination and/or composition is provided
in one or more nanoparticles. [0338] 51. The method of any of the
preceding paragraphs, wherein the combination is provided in the
form of one or more nanoparticles comprising the active compound,
the nanoparticles in solution or suspension in a composition
comprising the ionic liquid. [0339] 52. A composition comprising at
least one ionic liquid comprising: [0340] a hydrophobic anion
comprising a carboxylic acid having a pKa of at least 4.0 and a Log
P of at least 1.0; and [0341] a cation comprising a quaternary
ammonium. [0342] 53. The composition of paragraph 52, wherein the
anion has a pKa of at least 4.5. [0343] 54. The composition of any
of paragraphs 52-53, wherein the anion has a pKa of at least 5.0.
[0344] 55. The composition of any of paragraphs 52-54, wherein the
anion has a Log P of at least 2.0. [0345] 56. The composition of
any of paragraphs 52-55, wherein the anion has a Log P of at least
2.5. [0346] 57. The composition of any of paragraphs 52-56, wherein
the anion has a Log P of at least 2.75. [0347] 58. The composition
of any of paragraphs 52-57, wherein the anion comprises a carbon
chain of at least 8 carbons. [0348] 59. The composition of any of
paragraphs 52-58, wherein the anion is an alkene. [0349] 60. The
composition of any of paragraphs 52-59, wherein the anion is
geranic acid, octanoic acid, or citronellic acid. [0350] 61. The
composition of any of paragraphs 52-60, wherein the cation has a
molar mass equal to or greater than choline. [0351] 62. The
composition of any of paragraphs 52-61, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and at least one R
group comprises a hydroxy group. [0352] 63. The composition of any
of paragraphs 52-62, wherein the quarternary ammonium has the
structure of NR.sub.4.sup.+ and only one R group comprises a
hydroxy group. [0353] 64. The composition of any of paragraphs
52-63, wherein the cation is C1, C6, or C7. [0354] 65. The
composition of any of paragraphs 52-64, wherein the cation is
selected from choline, C1, C6, and C7 and the anion is citronellic
acid. [0355] 66. The composition of any of paragraphs 52-65,
wherein the cation is C1 and the anion is citronellic acid. [0356]
67. The composition of any of paragraphs 52-66, wherein the cation
is selected from C1, C6, and C7 and the anion is geranic acid.
[0357] 68. The composition of any of paragraphs 52-67, wherein the
ionic liquid is choline: citronellic acid, C1: geranic acid, or C1:
citronellic acid. [0358] 69. The composition of any of paragraphs
52-68, wherein the ionic liquid is not CAGE. [0359] 70. The
composition of any of paragraphs 52-69, wherein the ionic liquid
comprises a ratio of cation to anion of from about 2:1 to about
1:10. [0360] 71. The composition of any of paragraphs 52-70,
wherein the ionic liquid comprises a ratio of cation to anion of
from about 1:1 to about 1:4. [0361] 72. The composition of any of
paragraphs 52-71, wherein the ionic liquid comprises a ratio of
cation to anion of about 1:2. [0362] 73. The composition of any of
paragraphs 52-72, wherein the ionic liquid has a cation:anion ratio
of less than 1:1. [0363] 74. The composition of any of paragraphs
52-73, wherein the ionic liquid has a cation:anion ratio with an
excess of anion. [0364] 75. The composition of any of paragraphs
52-74, wherein the ionic liquid has less than 20 cross peaks as
measured by Nuclear Overhauser Effect SpectroscopY (NOESY). [0365]
76. The composition of any of paragraphs 52-75, wherein the ionic
liquid has less than 10 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY). [0366] 77. The composition
of any of paragraphs 52-76, wherein the ionic liquid has less than
5 cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY). [0367] 78. The composition of any of paragraphs 52-77,
further comprising at least one active compound in combination with
the at least one ionic liquid. [0368] 79. The composition of
paragraph 78, wherein the active compound is hydrophobic. [0369]
80. The composition of any of paragraphs 78-79, wherein the active
compound is hydrophilic. [0370] 81. The composition of any of
paragraphs 78-80, wherein the active compound comprises a
polypeptide. [0371] 82. The composition of any of paragraphs 78-81,
wherein the active compound has a molecular weight of greater than
450. [0372] 83. The composition of any of paragraphs 78-82, wherein
the active compound has a molecular weight of greater than 500.
[0373] 84. The composition of any of paragraphs 78-83, wherein the
active compound comprises an antibody or antibody reagent. [0374]
85. The composition of any of paragraphs 78-84, wherein the active
compound comprises insulin, acarbose, ruxolitinib, or a GLP-1
polypeptide or mimetic or analog thereof. [0375] 86. The
composition of any of paragraphs 52-85, wherein the ionic liquid is
at a concentration of at least 0.1% w/v. [0376] 87. The composition
of any of paragraphs 52-86, wherein the ionic liquid is at a
concentration of from about 10 to about 70% w/v. [0377] 88. The
composition of any of paragraphs 52-87, wherein the ionic liquid is
at a concentration of from about 30 to about 50% w/v. [0378] 89.
The composition of any of paragraphs 52-88, wherein the ionic
liquid is at a concentration of from about 30 to about 40% w/v.
[0379] 90. The composition of any of paragraphs 52-89, wherein the
compostition is formulated for transdermal administration. [0380]
91. The composition of any of paragraphs 52-90, wherein the
composition is formulated for administration transdermally, to a
mucus membrane, orally, subcutaneously, intradermally,
parenterally, intratumorally, or intravenously. [0381] 92. The
composition of paragraph 91, wherein the mucus membrane is nasal,
oral, or vaginal. [0382] 93. The composition of any of paragraphs
78-92, wherein the active compound is provided at a dosage of 1-20
mg/kg. [0383] 94. The composition of any of paragraphs 52-93,
further comprising at least one non-ionic surfactant. [0384] 95.
The composition of any of paragraphs 52-94, further comprising a
pharmaceutically acceptable carrier. [0385] 96. The composition of
any of paragraphs 52-95, wherein the composition is provided in a
degradable capsule. [0386] 97. The composition of any of paragraphs
78-96, wherein the composition is an admixture. [0387] 98. The
composition of any of paragraphs 52-97, wherein the composition is
provided in one or more nanoparticles. [0388] 99. The composition
of any of paragraphs 52-98, comprising one or more nanoparticles
comprising the active compound, the nanoparticles in solution or
suspension in a composition comprising the ionic liquid.
[0389] Some embodiments of the technology described herein can be
defined according to any of the following numbered paragraphs:
[0390] 1. A method of administering at least one active compound,
the method comprising administering the active compound in
combination with at least one ionic liquid comprising: [0391] a
hydrophobic anion comprising a carboxylic acid having a pKa of at
least 4.0 and a Log P of at least 1.0; and [0392] a cation
comprising a quaternary ammonium. [0393] 2. A method of reducing
weight/weight gain or treating obesity, diabetes, ulcers, cancer,
or fibrosis in a subject in need thereof, the method comprising
administering a composition comprising at least one ionic liquid
comprising: [0394] a hydrophobic anion comprising a carboxylic acid
having a pKa of at least 4.0 and a Log P of at least 1.0; and
[0395] a cation comprising a quaternary ammonium to the subject.
[0396] 3. The method of paragraph 2, wherein the composition does
not comprise a therapeutic agent other than the at least one ionic
liquid. [0397] 4. The method of paragraph 2, wherein the
composition further comprises an active compound other than the at
least one ionic liquid. [0398] 5. The method of any of the
preceding paragraphs, wherein the anion has a pKa of at least 4.5.
[0399] 6. The method of any of the preceding paragraphs, wherein
the anion has a pKa of at least 5.0. [0400] 7. The method of any of
the preceding paragraphs, wherein the anion has a pKa of at least
4.895. [0401] 8. The method of any of the preceding paragraphs,
wherein the anion has a pKa of 4.5-5.5. [0402] 9. The method of any
of the preceding paragraphs, wherein the anion has a pKa of
4.895-5.19. [0403] 10. The method of any of the preceding
paragraphs, wherein the anion has a Log P of at least 2.0. [0404]
11. The method of any of the preceding paragraphs, wherein the
anion has a Log P of at least 2.5. [0405] 12. The method of any of
the preceding paragraphs, wherein the anion has a Log P of at least
2.75. [0406] 13. The method of any of the preceding paragraphs,
wherein the anion has a Log P of at least 2.8. [0407] 14. The
method of any of the preceding paragraphs, wherein the anion has a
Log P of 2.5-3.5. [0408] 15. The method of any of the preceding
paragraphs, wherein the anion has a Log P of 2.8-3.01. [0409] 16.
The method of any of the preceding paragraphs, wherein the anion
comprises a carbon chain of at least 8 carbons. [0410] 17. The
method of any of the preceding paragraphs, wherein the anion
comprises a carbon chain with an 8 carbon backbone. [0411] 18. The
method of any of the preceding paragraphs, wherein the anion is
geranic acid, octenoic acid, octanoic acid, or citronellic acid.
[0412] 19. The method of any of the preceding paragraphs, wherein
the anion is octenoic acid, octanoic acid, or citronellic acid.
[0413] 20. The method of any of the preceding paragraphs, wherein
the anion is an alkene. [0414] 21. The method of any of the
preceding paragraphs, wherein the anion is geranic acid, octanoic
acid, or citronellic acid. [0415] 22. The method of any of the
preceding paragraphs, wherein the cation has a molar mass equal to
or greater than choline. [0416] 23. The method of any of the
preceding paragraphs, wherein the quarternary ammonium has the
structure of NR.sub.4.sup.+ and at least one R group comprises a
hydroxy group. [0417] 24. The method of any of the preceding
paragraphs, wherein the quarternary ammonium has the structure of
NR.sub.4.sup.+ and only one R group comprises a hydroxy group.
[0418] 25. The method of any of the preceding paragraphs, wherein
the cation is C1, C6, or C7. [0419] 26. The method of any of the
preceding paragraphs, wherein the cation is selected from choline,
C1, C6, and C7 and the anion is citronellic acid. [0420] 27. The
method of any of the preceding paragraphs, wherein the cation is C1
and the anion is citronellic acid. [0421] 28. The method of any of
the preceding paragraphs, wherein the cation is selected from C1,
C6, and C7 and the anion is geranic acid. [0422] 29. The method of
any of the preceding paragraphs, wherein the ionic liquid is
choline: citronellic acid, C1: geranic acid, or C1: citronellic
acid. [0423] 30. The method of any of paragraphs 1-16, wherein the
cation is selected from choline, C1, C6, and C7 and the anion is
selected from citronellic acid, octanoic acid, and octenoic acid.
[0424] 31. The method of any of paragraphs 1-16, wherein the cation
is choline and the anion is selected from citronellic acid,
octanoic acid, and octenoic acid. [0425] 32. The method of any of
paragraphs 1-16, wherein the ionic liquid is choline: citronellic
acid, choline: octanoic acid, or choline: octenoic acid. [0426] 33.
The method of any of the preceding paragraphs, wherein the ionic
liquid is not CAGE. [0427] 34. The method of any of the preceding
paragraphs, wherein the ionic liquid has less than 20 cross peaks
as measured by Nuclear Overhauser Effect SpectroscopY (NOESY).
[0428] 35. The method of any of the preceding paragraphs, wherein
the ionic liquid has less than 10 cross peaks as measured by
Nuclear Overhauser Effect SpectroscopY (NOESY). [0429] 36. The
method of any of the preceding paragraphs, wherein the ionic liquid
has less than 5 cross peaks as measured by Nuclear Overhauser
Effect SpectroscopY (NOESY). [0430] 37. The method of any of the
preceding paragraphs, wherein the administration is transdermal.
[0431] 38. The method of any of the preceding paragraphs, wherein
the administration is transdermal, to a mucus membrane, oral,
subcutaneous, intradermal, parenteral, intratumoral, or
intravenous. [0432] 39. The method of paragraph 26, wherein the
mucus membrane is nasal, oral, or vaginal. [0433] 40. The method of
any of the preceding paragraphs, wherein the administration is
oral. [0434] 41. The method of any of the preceding paragraphs,
wherein the ionic liquid is at a concentration of at least 0.1%
w/v. [0435] 42. The method of any of the preceding paragraphs,
wherein the ionic liquid is at a concentration of from about 10 to
about 70% w/v. [0436] 43. The method of any of the preceding
paragraphs, wherein the ionic liquid is at a concentration of from
about 30 to about 50% w/v. [0437] 44. The method of any of the
preceding paragraphs, wherein the ionic liquid is at a
concentration of from about 30 to about 40% w/v. [0438] 45. The
method of any of the preceding paragraphs, wherein the ionic liquid
comprises a ratio of cation to anion of from about 2:1 to about
1:10. [0439] 46. The method of any of the preceding paragraphs,
wherein the ionic liquid comprises a ratio of cation to anion of
from about 1:1 to about 1:4. [0440] 47. The method of any of the
preceding paragraphs, wherein the ionic liquid comprises a ratio of
cation to anion of about 1:2. [0441] 48. The method of any of the
preceding paragraphs, wherein the ionic liquid has a cation:anion
ratio of less than 1:1. [0442] 49. The method of any of the
preceding paragraphs, wherein the active compound is hydrophobic.
[0443] 50. The method of any of the preceding paragraphs, wherein
the active compound is hydrophilic. [0444] 51. The method of any of
the preceding paragraphs, wherein the active compound comprises a
polypeptide. [0445] 52. The method of any of the preceding
paragraphs, wherein the active compound has a molecular weight of
greater than 450. [0446] 53. The method of any of the preceding
paragraphs, wherein the active compound has a molecular weight of
greater than 500. [0447] 54. The method of any of the preceding
paragraphs, wherein the active compound comprises an antibody or
antibody reagent. [0448] 55. The method of any of the preceding
paragraphs, wherein the active compound comprises insulin,
acarbose, ruxolitinib, or a GLP-1 polypeptide or mimetic or analog
thereof [0449] 56. The method of any of the preceding paragraphs,
wherein the combination and/or composition is administered once.
[0450] 57. The method of any of the preceding paragraphs, wherein
the combination and/or composition is administered in multiple
doses. [0451] 58. The method of any of the preceding paragraphs,
wherein the active compound and/or composition is provided at a
dosage of 1-20 mg/kg. [0452] 59. The method of any of the preceding
paragraphs, wherein the active compound and the ionic liquid are
further in combination with at least one non-ionic surfactant.
[0453] 60. The method of any of the preceding paragraphs, wherein
the combination and/or composition further comprises a further
pharmaceutically acceptable carrier. [0454] 61. The method of any
of the preceding paragraphs, wherein the administration is oral and
the combination and/or composition is provided in a degradable
capsule. [0455] 62. The method of any of the preceding paragraphs,
wherein the combination is an admixture. [0456] 63. The method of
any of the preceding paragraphs, wherein the combination and/or
composition is provided in one or more nanoparticles. [0457] 64.
The method of any of the preceding paragraphs, wherein the
combination is provided in the form of one or more nanoparticles
comprising the active compound, the nanoparticles in solution or
suspension in a composition comprising the ionic liquid. [0458] 65.
A composition comprising at least one ionic liquid comprising:
[0459] a hydrophobic anion comprising a carboxylic acid having a
pKa of at least 4.0 and a Log P of at least 1.0; and [0460] a
cation comprising a quaternary ammonium. [0461] 66. The composition
of paragraph 65, wherein the anion has a pKa of at least 4.5.
[0462] 67. The composition of paragraph 65, wherein the anion has a
pKa of at least 4.895. [0463] 68. The composition of paragraph 65,
wherein the anion has a pKa of 4.5-5.5. [0464] 69. The composition
of paragraph 65, wherein the anion has a pKa of 4.895-5.19. [0465]
70. The composition of any of paragraphs 65-69, wherein the anion
has a pKa of at least 5.0. [0466] 71. The composition of any of
paragraphs 65-69, wherein the anion has a Log P of at least 2.0.
[0467] 72. The composition of any of paragraphs 65-69, wherein the
anion has a Log P of at least 2.5. [0468] 73. The composition of
any of paragraphs 65-69, wherein the anion has a Log P of at least
2.75. [0469] 74. The composition of any of paragraphs 65-69,
wherein the anion has a Log P of at least 2.8. [0470] 75. The
composition of any of paragraphs 65-69, wherein the anion has a Log
P of 2.5-3.5. [0471] 76. The composition of any of paragraphs
65-69, wherein the anion has a Log P of 2.8-3.01. [0472] 77. The
composition of any of paragraphs 65-76, wherein the anion comprises
a carbon chain of at least 8 carbons. [0473] 78. The composition of
any of paragraphs 65-76, wherein the anion comprises a carbon chain
with an 8 carbon backbone. [0474] 79. The composition of any of
paragraphs 65-76, wherein the anion is geranic acid, octenoic acid,
octanoic acid, or citronellic acid. [0475] 80. The composition of
any of paragraphs 65-76, wherein the anion is octenoic acid,
octanoic acid, or citronellic acid. [0476] 81. The composition of
any of paragraphs 65-80, wherein the anion is an alkene. [0477] 82.
The composition of any of paragraphs 65-81, wherein the anion is
geranic acid, octanoic acid, or citronellic acid. [0478] 83. The
composition of any of paragraphs 65-82, wherein the cation has a
molar mass equal to or greater than choline. [0479] 84. The
composition of any of paragraphs 65-83, wherein the quarternary
ammonium has the structure of NR.sub.4.sup.+ and at least one R
group comprises a hydroxy group. [0480] 85. The composition of any
of paragraphs 65-84, wherein the quarternary ammonium has the
structure of NR.sub.4.sup.+ and only one R group comprises a
hydroxy group. [0481] 86. The composition of any of paragraphs
65-85, wherein the cation is C1, C6, or C7. [0482] 87. The
composition of any of paragraphs 65-86, wherein the cation is
selected from choline, C1, C6, and C7 and the anion is citronellic
acid. [0483] 88. The composition of any of paragraphs 65-87,
wherein the cation is C1 and the anion is citronellic acid. [0484]
89. The composition of any of paragraphs 65-88, wherein the cation
is selected from C1, C6, and C7 and the anion is geranic acid.
[0485] 90. The composition of any of paragraphs 65-89, wherein the
ionic liquid is choline: citronellic acid, C1: geranic acid, or C1:
citronellic acid. [0486] 91. The composition of any of paragraphs
65-90, wherein the cation is selected from choline, C1, C6, and C7
and the anion is selected from citronellic acid, octanoic acid, and
octenoic acid. [0487] 92. The composition of any of paragraphs
65-91, wherein the cation is choline and the anion is selected from
citronellic acid, octanoic acid, and octenoic acid. [0488] 93. The
composition of any of paragraphs 65-92, wherein the ionic liquid is
choline: citronellic acid, choline: octanoic acid, or choline:
octenoic acid. [0489] 94. The composition of any of paragraphs
65-93, wherein the ionic liquid is not CAGE. [0490] 95. The
composition of any of paragraphs 65-94, wherein the ionic liquid
comprises a ratio of cation to anion of from about 2:1 to about
1:10. [0491] 96. The composition of any of paragraphs 65-95,
wherein the ionic liquid comprises a ratio of cation to anion of
from about 1:1 to about 1:4. [0492] 97. The composition of any of
paragraphs 65-96, wherein the ionic liquid comprises a ratio of
cation to anion of about 1:2. [0493] 98. The composition of any of
paragraphs 65-97, wherein the ionic liquid has a cation:anion ratio
of less than 1:1. [0494] 99. The composition of any of paragraphs
65-98, wherein the ionic liquid has a cation:anion ratio with an
excess of anion. [0495] 100. The composition of any of paragraphs
65-99, wherein the ionic liquid has less than 20 cross peaks as
measured by Nuclear Overhauser Effect SpectroscopY (NOESY). [0496]
101. The composition of any of paragraphs 65-100, wherein the ionic
liquid has less than 10 cross peaks as measured by Nuclear
Overhauser Effect SpectroscopY (NOESY). [0497] 102. The composition
of any of paragraphs 65-101, wherein the ionic liquid has less than
5 cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY). [0498] 103. The composition of any of paragraphs 65-102,
further comprising at least one active compound in combination with
the at least one ionic liquid. [0499] 104. The composition of
paragraph 103, wherein the active compound is hydrophobic. [0500]
105. The composition of any of paragraphs 103-104, wherein the
active compound is hydrophilic. [0501] 106. The composition of any
of paragraphs 103-105, wherein the active compound comprises a
polypeptide. [0502] 107. The composition of any of paragraphs
103-106, wherein the active compound has a molecular weight of
greater than 450. [0503] 108. The composition of any of paragraphs
103-107, wherein the active compound has a molecular weight of
greater than 500. [0504] 109. The composition of any of paragraphs
103-108, wherein the active compound comprises an antibody or
antibody reagent.
[0505] 110. The composition of any of paragraphs 103-109, wherein
the active compound comprises insulin, acarbose, ruxolitinib, or a
GLP-1 polypeptide or mimetic or analog thereof [0506] 111. The
composition of any of paragraphs 65-110, wherein the ionic liquid
is at a concentration of at least 0.1% w/v. [0507] 112. The
composition of any of paragraphs 65-111, wherein the ionic liquid
is at a concentration of from about 10 to about 70% w/v. [0508]
113. The composition of any of paragraphs 65-112, wherein the ionic
liquid is at a concentration of from about 30 to about 50% w/v.
[0509] 114. The composition of any of paragraphs 65-113, wherein
the ionic liquid is at a concentration of from about 30 to about
40% w/v. [0510] 115. The composition of any of paragraphs 65-114,
wherein the composition is formulated for transdermal
administration. [0511] 116. The composition of any of paragraphs
65-115, wherein the composition is formulated for administration
transdermally, to a mucus membrane, orally, subcutaneously,
intradermally, parenterally, intratumorally, or intravenously.
[0512] 117. The composition of paragraph 116, wherein the mucus
membrane is nasal, oral, or vaginal. [0513] 118. The composition of
any of paragraphs 65-117, wherein the composition is formulated for
oral administration. [0514] 119. The composition of any of
paragraphs 103-118, wherein the active compound is provided at a
dosage of 1-20 mg/kg. [0515] 120. The composition of any of
paragraphs 65-119, further comprising at least one non-ionic
surfactant. [0516] 121. The composition of any of paragraphs
65-120, further comprising a pharmaceutically acceptable carrier.
[0517] 122. The composition of any of paragraphs 65-121, wherein
the composition is provided in a degradable capsule. [0518] 123.
The composition of any of paragraphs 65-122, wherein the
composition is an admixture. [0519] 124. The composition of any of
paragraphs 65-123, wherein the composition is provided in one or
more nanoparticles. [0520] 125. The composition of any of
paragraphs 65-124, comprising one or more nanoparticles comprising
the active compound, the nanoparticles in solution or suspension in
a composition comprising the ionic liquid. [0521] 126. A method of
designing and/or identifying an ionic liquid comprising two ions,
wherein one ion is a cation and the other ion is an anion, the
method comprising: [0522] a. selecting one of the two ions of the
ionic liquid; and [0523] b. selecting the other ion to minimize
inter-ionic interactions. [0524] 127. A method of designing and/or
identifying an ionic liquid comprising two ions, wherein one ion is
a cation and the other ion is an anion, the method comprising:
[0525] a. selecting the cation; and [0526] b. selecting the anion
to minimize inter-ionic interactions. [0527] 128. A method of
designing and/or identifying an ionic liquid comprising two ions,
wherein one ion is a cation and the other ion is an anion, the
method comprising: [0528] a. selecting the anion; and [0529] b.
selecting the cation to minimize inter-ionic interactions. [0530]
129. A method of designing and/or identifying an ionic liquid
comprising two ions, wherein one ion is a cation and the other ion
is an anion, from a pool of candidate cations and a pool of
candidate anions, the method comprising: [0531] a. selecting one of
the two ions of the ionic liquid from the pool of candidate ions;
and [0532] b. selecting from the other pool of candidate ions the
ion which most minimizes inter-ionic interactions with the ion
selected in step a. [0533] 130. A method of designing and/or
identifying an ionic liquid comprising two ions, wherein one ion is
a cation and the other ion is an anion, from a pool of candidate
cations and a pool of candidate anions, the method comprising:
[0534] a. selecting the cation from the pool of candidate cations;
[0535] b. selecting from the pool of candidate anions the anion
which most minimizes inter-ionic interactions with the cation
selected in step a. [0536] 131. A method of designing and/or
identifying an ionic liquid comprising two ions, wherein one ion is
a cation and the other ion is an anion, from a pool of candidate
cations and a pool of candidate anions, the method comprising:
[0537] a. selecting the cation from the pool of candidate anions;
[0538] b. selecting from the pool of candidate cations the anion
which most minimizes inter-ionic interactions with the anion
selected in step a. [0539] 132. The method of any of paragraphs
126-131, wherein the ionic liquid is selected or designed for
transdermal administration. [0540] 133. The method of any of
paragraphs 126-132, wherein the ionic liquid is selected or
designed for administration transdermally, to a mucus membrane,
orally, subcutaneously, intradermally, parenterally,
intratumorally, or intravenously. [0541] 134. The method of
paragraph 133, wherein the mucus membrane is nasal, oral, or
vaginal. [0542] 135. The method of any of paragraphs 126-134,
wherein the ionic liquid is selected or designed for oral
administration. [0543] 136. The method of any of paragraphs
126-135, wherein the ionic liquid is selected or designed for
delivery of an active compound. [0544] 137. The method of any of
paragraphs 126-136, wherein the cation comprises, or selecting the
cation comprises selection a cation that comprises a quaternary
ammonium; and the anion comprises or selecting an anion comprises
selecting a hydrophobic anion comprising a carboxylic acid having a
pKa of at least 4.0 and a Log P of at least 1.0. [0545] 138. The
method of any of paragraphs 126-137, wherein the anion has, or
selecting an anion comprises selecting an anion that has, a pKa of
at least 4.5. [0546] 139. The method of any of paragraphs 126-138,
wherein the anion has, or selecting an anion comprises selecting an
anion that has, a pKa of at least 4.895. [0547] 140. The method of
any of paragraphs 126-138, wherein the anion has, or selecting an
anion comprises selecting an anion that has, a pKa of 4.5-5.5.
[0548] 141. The method of any of paragraphs 126-140, wherein the
anion has, or selecting an anion comprises selecting an anion that
has, a pKa of 4.895-5.19. [0549] 142. The method of any of
paragraphs 126-141, wherein the anion has, or selecting an anion
comprises selecting an anion that has, a pKa of at least 5.0.
[0550] 143. The method of any of paragraphs 126-142, wherein the
anion has, or selecting an anion comprises selecting an anion that
has, a Log P of at least 2.0. [0551] 144. The method of any of
paragraphs 126-143, wherein the anion has, or selecting an anion
comprises selecting an anion that has, a Log P of at least 2.5.
[0552] 145. The method of any of paragraphs 126-144, wherein the
anion has, or selecting an anion comprises selecting an anion that
has, a Log P of at least 2.75. [0553] 146. The method of any of
paragraphs 126-145, wherein the anion has, or selecting an anion
comprises selecting an anion that has, a Log P of at least 2.8.
[0554] 147. The method of any of paragraphs 126-146, wherein the
anion has, or selecting an anion comprises selecting an anion that
has, a Log P of 2.5-3.5. [0555] 148. The method of any of
paragraphs 126-147, wherein the anion has, or selecting an anion
comprises selecting an anion that has, a Log P of 2.8-3.01. [0556]
149. The method of any of paragraphs 126-148, wherein the anion
comprises, or selecting an anion comprises selecting an anion that
comprises, a carbon chain of at least 8 carbons. [0557] 150. The
method of any of paragraphs 126-149, wherein the anion comprises,
or selecting an anion comprises selecting an anion that comprises,
a carbon chain with an 8 carbon backbone. [0558] 151. The method of
any of paragraphs 126-150, wherein the anion is, or selecting an
anion comprises selecting an anion that is, geranic acid, octenoic
acid, octanoic acid, or citronellic acid. [0559] 152. The method of
any of paragraphs 126-151, wherein the anion is, or selecting an
anion comprises selecting an anion that is, octenoic acid, octanoic
acid, or citronellic acid. [0560] 153. The method of any of
paragraphs 126-152, wherein the anion is, or selecting an anion
comprises selecting an anion that is, an alkene. [0561] 154. The
method of any of paragraphs 126-153, wherein the anion is, or
selecting an anion comprises selecting an anion that is, geranic
acid, octanoic acid, or citronellic acid. [0562] 155. The method of
any of paragraphs 126-154, wherein the cation has, or selecting a
cation comprises selecting a cation that has, a molar mass equal to
or greater than choline. [0563] 156. The method of any of
paragraphs 126-155, wherein the quarternary ammonium has the
structure of NR.sub.4+ and at least one R group comprises a hydroxy
group. [0564] 157. The method of any of paragraphs 126-156, wherein
the quarternary ammonium has the structure of NR.sub.4+ and only
one R group comprises a hydroxy group. [0565] 158. The method of
any of paragraphs 126-157, wherein the cation is, or selecting a
cation comprises selecting a cation that is, C1, C6, or C7. [0566]
159. The method of any of paragraphs 126-158, wherein the cation
is, or selecting a cation comprises selecting a cation that is,
selected from choline, C1, C6, and C7 and the anion is citronellic
acid. [0567] 160. The method of any of paragraphs 126-159, wherein
the cation is, or selecting a cation comprises selecting a cation
that is, selected from C1, C6, and C7 and the anion is geranic
acid. [0568] 161. The method of any of paragraphs 126-160, wherein
the cation is, or selecting a cation comprises selecting a cation
that is, choline, C1, C6, or C7 and selecting the anion comprises
selecting an anion that is citronellic acid, octanoic acid, or
octenoic acid. [0569] 162. The method of any of paragraphs 126-161,
wherein the cation is choline and selecting the anion comprises
selecting an anion that is citronellic acid, octanoic acid, or
octenoic acid. [0570] 163. The method of any of paragraphs 126-162,
wherein the ionic liquid is not CAGE. [0571] 164. The method of any
of paragraphs 126-163, wherein the ionic liquid comprises a ratio
of cation to anion of from about 2:1 to about 1:10. [0572] 165. The
method of any of paragraphs 126-164, wherein the ionic liquid
comprises a ratio of cation to anion of from about 1:1 to about
1:4. [0573] 166. The method of any of paragraphs 126-165, wherein
the ionic liquid comprises a ratio of cation to anion of about 1:2.
[0574] 167. The method of any of paragraphs 126-166, wherein the
ionic liquid has a cation:anion ratio of less than 1:1. [0575] 168.
The method of any of paragraphs 126-167, wherein the ionic liquid
has a cation:anion ratio with an excess of anion. [0576] 169. The
method of any of paragraphs 126-168, wherein minimizing inter-ionic
interaction comprises minimizing the number of cross peaks as
measured by Nuclear Overhauser Effect SpectroscopY (NOESY). [0577]
170. The method of any of paragraphs 126-169, wherein a cation and
anion minimize inter-ionic interaction if they have less than 20
cross peaks as measured by Nuclear Overhauser Effect SpectroscopY
(NOESY). [0578] 171. The method of any of paragraphs 126-170,
wherein a cation and anion minimize inter-ionic interaction if they
have less than 10 cross peaks as measured by Nuclear Overhauser
Effect SpectroscopY (NOESY). [0579] 172. The method of any of
paragraphs 126-171, wherein a cation and anion minimize inter-ionic
interaction if they have less than 5 cross peaks as measured by
Nuclear Overhauser Effect SpectroscopY (NOESY). [0580] 173. The
method of any of paragraphs 126-172, wherein the active compound is
hydrophobic. [0581] 174. The method of any of paragraphs 126-173,
wherein the active compound is hydrophilic. [0582] 175. The method
of any of paragraphs 126-174, wherein the active compound comprises
a polypeptide. [0583] 176. The method of any of paragraphs 126-175,
wherein the active compound has a molecular weight of greater than
450. [0584] 177. The method of any of paragraphs 126-176, wherein
the active compound has a molecular weight of greater than 500.
[0585] 178. The method of any of paragraphs 126-177, wherein the
active compound comprises an antibody or antibody reagent. [0586]
179. The method of any of paragraphs 126-178, wherein the active
compound comprises insulin, acarbose, ruxolitinib, or a GLP-1
polypeptide or mimetic or analog thereof [0587] 180. The method of
any of paragraphs 126-179, wherein the ionic liquid is at a
concentration of at least 0.1% w/v. [0588] 181. The method of any
of paragraphs 126-180, wherein the ionic liquid is at a
concentration of from about 10 to about 70% w/v. [0589] 182. The
method of any of paragraphs 126-181, wherein the ionic liquid is at
a concentration of from about 30 to about 50% w/v. [0590] 183. The
method of any of paragraphs 126-182, wherein the ionic liquid is at
a concentration of from about 30 to about 40% w/v. [0591] 184. The
method of any of paragraphs 126-183, wherein the active compound is
provided at a dosage of 1-20 mg/kg. [0592] 185. The method of any
of paragraphs 126-184, wherein the ionic liquid is designed or
selected to be provided in a degradable capsule. [0593] 186. The
method of any of paragraphs 126-185, wherein the ionic liquid and
active compound are in admixture. [0594] 187. The method of any of
paragraphs 126-186, wherein the ionic liquid and optionally the
active compound are selected or designed to be provided in one or
more nanoparticles. [0595] 188. The method of any of paragraphs
126-187, wherein the ionic liquid is in solution or suspension in a
composition with one or more nanoparticles comprising the active
compound.
EXAMPLES
Example 1
[0596] Choline-based ionic liquids, in particular choline and
geranic acid (CAGE), have been used to enhance the delivery of
several small and large molecules across the skin. However,
detailed studies outlining the design principles of ILs for
transdermal drug delivery are still lacking. Using two model drugs
of differing hydrophilicities, acarbose and ruxolitinib, and 16
ionic liquids of varying cations and anions, we examine herein the
dependence of skin penetration on the chemical properties of ILs.
First, the impact of ion stoichiometry on skin penetration of drugs
was assessed using CAGE as a representative IL, and it showed that
a molar ratio of 1:2 of choline to geranic acid in CAGE yielded the
highest delivery. Subsequently, variants of CAGE were prepared
using anions with structural similarity to geranic acid and cations
with structural similarity to choline. The cation to anion ratio
was held constant at 1:2 for all variants. A range of permeation
enhancement effects was observed among the CAGE variants with some
variants outperforming the original CAGE composition. Mechanistic
studies revealed that the potency of ILs in enhancing transdermal
drug delivery correlated inversely with the inter-ionic
interactions as determined by 2D NMR. Using this understanding, a
new CAGE variant was designed, and it provided the highest delivery
of ruxolitinib of all ILs tested here. Overall, these studies
provide a generalized framework for optimizing ILs for enhancing
skin permeability.
Introduction
[0597] Transdermal drug delivery offers a painless mode of
administering drugs while concurrently avoiding first-pass
metabolism. [1] However, overcoming the uppermost transport barrier
of skin, the stratum corneum (SC), is a significant challenge.[1]
The SC consists of a tightly packed brick-and-mortar-like structure
of corneocytes, surrounded by lipids [2]. Only few drugs possessing
low mass and high lipophilicity are capable of navigating the SC
barrier without assistance. Several methods have been engineered to
enhance skin permeability to drugs including the use of ultrasound,
[3,4] microneedles, [5,6] iontophoresis, [7,8] jet injectors,
[9,10] and permeation enhancers, [11-13] among others. Recently,
ionic liquids and deep eutectic solvents (hereafter collectively
referred to as ionic liquids (ILs) for simplicity) have shown great
promise for transdermal drug delivery.[14-19] We have previously
demonstrated that a choline and geranic acid based IL, referred to
as CAGE (FIG. 1), has shown to be effective in transdermal delivery
of insulin [20] as well as small molecule antimicrobial
agents.[17]
[0598] The efficacy of ILs in enhancing skin permeability is
expected to depend on their ionic constituents and composition.
However, such knowledge-base is critically missing from the
literature. One study has shown that the efficacy of CAGE in
insulin delivery depends strongly on its ion stoichiometry. The
study further demonstrated that CAGE mediates its enhancement
effect primarily through its interaction of SC lipids.[16] However,
the generalized understanding of the role of ion properties in
determining IL's delivery efficacy is still lacking.
[0599] The work presented herein represents the first systematic
study on the role of anion and cation properties in IL-mediated
transdermal drug delivery. Drugs of differing hydrophobicities were
used to assess the transport potential of the ILs (FIG. 2). The
first drug, acarbose, is a hydrophilic molecule used to treat Type
2 diabetes by inhibiting enzymes that digest carbohydrates. [27] It
is currently administered orally but has a very low bioavailability
and a high gastrointestinal side effect profile and therefore has
limited use. [28] The second drug, ruxolitinib is a hydrophobic
janus kinase (JAK) inhibitor that is currently FDA approved as an
oral pill for use in myelofibrosis. [29,30] It has also shown
promise in treating alopecia, therefore its topical delivery is of
interest. [31,32] The ILs used in this study were built around the
composition of CAGE. Specifically, eight organic acids of varying
molecular weights, pKas and hydrophobicities were used as
alternatives to geranic acid, and seven organic quaternary amines
were used as alternatives to cholines. The resultant ILs were
characterized by NMR and their ability to deliver ruxolitinib and
acarbose was measured in vitro.
[0600] Methods and Materials
[0601] Materials
[0602] Choline bicarbonate, geranic acid, the tertiary amines,
halogenated alcohols, Phosphate Buffered Saline (PBS), D20 and
deuterated DMSO were obtained from Sigma Aldrich (St. Louis, Mo.,
USA). The commercial geranic acid (85%, yellow liquid) was purified
prior to use with at least five successive freeze-thaw cycles with
acetone until the solution was clear. The residual acetone was
removed under reduced pressure prior to use. Porcine skin (Lampire
Biological Laboratories, Pipersville, Pa.) was kept at -80'C and
thawed immediately prior to use.
[0603] Methods
[0604] Synthesis of CAGE. CAGE was synthesized at various ion
stoichiometries as previously reported. [17] Choline bicarbonate
and geranic acid was mixed at various molar ratios including 1:4,
1:2, 1:1 and 2:1 to prepare CAGE by salt metathesis reaction. Each
CAGE composition was characterized via NMR with DMSO-d.sub.6 on an
Agilent DD2.TM. 600 MHz spectrometer.
[0605] Synthesis of Choline Analogs. Seven variants of choline were
prepared to assess the dependence of transport on cation
properties. The synthesis was modelled on that reported in Ref
[33]. The starting material for all variants was either a
triethylamine, tripropylamine or tributylamine. In each case, the
amine was reacted with a halogenated alcohol in a 1:1 molar ratio
by dissolution in toluene (30 mL) and heating in a round bottom
flask for 12 hours at a pre-determined temperature Table 4). The
various reactants, the reaction temperatures, and key information
about the resulting products can be found in Table 4. A majority of
the solvent was then removed using a rotary evaporator at 20 mbar
at 60.degree. C. for 2 hours. The residual solvent was removed
under reduced pressure at 60.degree. C. for 48 hours.
[0606] ILs of Geranic Acid with Choline Alternatives. Quaternary
amines were reacted with recrystallized geranic acid in a 1:2 molar
ratio at 40.degree. C. for 12 hours, similarly to the CAGE
synthesis reported previously.[34] A rotary evaporator was then
used to dry the resulting ionic liquids at 20 mbar at 60.degree. C.
for 2 hours. The residual water was removed under reduced pressure
at 60.degree. C. for 48 hours.
[0607] ILs with Geranic Acid Alternatives. Carboxylic acids
dissolved in the minimum amount of ultrapure water (or methanol in
the case of salicylic acid) needed for dissolution (<5 mL) were
reacted with choline bicarbonate in a 1:2 molar ratio (choline:
carboxylic acid) at 40.degree. C. for 12 hours. A rotary evaporator
was then used to dry the resulting ionic liquids at 20 mbar at
60.degree. C. for 2 hours. The residual water was removed under
reduced pressure at 60.degree. C. for 48 hours.
[0608] NOESY measurements. Samples were prepared by placing dried
IL into an NMR tube alongside a coaxial insert containing
deuterated DMSO. An Agilent DD2TM 600 MHz spectrometer was used. A
90.degree. pulse width of 11.25 was employed, number of increments
(ni) was set to 128 and the number of scans was set to 4. The
spectra were phase corrected after measurement.
[0609] Skin Penetration Studies. The skin penetration studies were
undertaken using porcine skin in Franz diffusion cells, as
described in full previously (skin area=1.7 cm.sup.2). [35]
Briefly, thawed, washed porcine skin was placed in a diffusion cell
with the SC facing upwards. The acceptor component of the cell was
filled with PBS and equipped with a magnetic stirrer bar. Only
cells with a measured trans-epidermal current less than 12 .mu.A
were used in the study and the treatments were randomly assigned.
300 .mu.L of drug solution in each IL (1 mg/mL) was placed on top
of the skin, ensuring full coverage. The cell was placed on a
stirrer plate at 37.degree. C. for 24 hours, at which point the
skin was removed and the surface was washed gently with PBS. Each
skin layer was then separated. The stratum corneum was removed by
tape stripping (up to ten layers), the epidermis was separated from
the dermis with a scalpel, and a 4 mm punch was used thrice to
remove a third of the dermis (by area). The tissue was then placed
into either methanol (ruxolitinib) or a 50% methanol/PBS mixture
(acarbose), and left to shake overnight to extract the drug, which
was then analyzed by HPLC, as described below.
[0610] HPLC Analysis. The analysis of the amount of drug present in
each layer was determined by HPLC. In each case a calibration curve
was prepared, consisting of ten samples covering a concentration
range of 2.times.10.sup.-3-1 mg/ml. For acarbose, a 250.times.4.6,
5 .mu.m Nucleosil-NH2 column was used. The isocratic mobile phase
was 1 mL/min acetonitrile:phosphate buffer (where 1 L contained 0.6
g of KH.sub.2PO.sub.4 and 0.48 g of Na.sub.2HPO.sub.4) in a ratio
of 76:24, and the UV detector (at 210 nm) showed a peak which
appeared at 5.2 minutes. For ruxolitinib, an Inertsil ODS C-18
column with 250.times.4.6 mm internal diameter and 5.mu.m particle
size was employed. A mobile phase of isocratically eluted THF:
Methanol: Acetonitrile 10:40:50 (v/v/v) with a flow rate of 1
mil/min was used, and the peak at 3.2 minutes was identified as the
product peak (detected by UV at 270 nm).
[0611] Results
[0612] Dependence of Delivery Efficacy on Ion Stoichiometry.
Initial studies were performed to assess the role of ion
stoichiometry in CAGE in determining its ability to deliver
acarbose and ruxolitinib. Four variants of CAGE were synthesized
with choline:geranic acid ratios of 2:1, 1:1, 1:2, and 1:4.
Acarbose and ruxolitinib were dissolved in CAGE at a concentration
of 1 mg/ml. Both drugs dissolved completely in all CAGE variants at
this concentration. Delivery of both drugs into skin exhibited a
strong dependence on CAGE composition (FIG. 3, acarbose (red) and
ruxolitinib (blue)). Amounts shown in FIG. 3 correspond those
delivered into dermis and acceptor compartment. Detailed
distribution of drugs in various skin layers is shown in FIGS. 11
and 12. Highest amounts of delivered drugs were found in the
dermis, followed by acceptor, followed by the epidermis. No
significant transport of either drug was observed from PBS
(negative control). Acarbose was soluble in PBS at a concentration
of 1 mg/ml, whereas ruxolitinib was soluble only at a concentration
of 0.2 mg/ml and hence a saturated solution in PBS was used in the
donor. In case of ruxolitinib, the delivery efficacy increased by
.about.4-fold as the geranic acid content increased by 2-fold (2:1
CAGE to 1:2 CAGE), after which it plateaued. A similar trend was
observed in case of acarbose, thus confirming that 1:2 CAGE
exhibits the highest delivery efficacy for hydrophilic as well as
lipophilic drugs. This is consistent with previous study with
insulin, whereby 1:2 CAGE performed the best out of the four
variants presented. [16] Note that no significant transport of
ruxolitinib or acarbose was observed when they were solubilized in
pure geranic acid or 80% choline bicarbonate in water. Given that
the 1:2 choline:geranic acid ratio provided maximum delivery
regardless of the drug hydrophilicity, subsequent variants were
prepared at this ion stoichiometry.
[0613] Dependence of Delivery Efficacy on Anion Chemistry. A first
set of IL variants was synthesized by replacing geranic acid with
other organic acids. Eight alternatives to geranic acid were used
(FIG. 4) The first, and the closest variant of geranic acid was
citronellic acid, which possesses one less double bond compared to
geranic acid and is otherwise identical. Octanoic acid possesses
the same number of carbons in the main backbone as geranic acid but
lacks unsaturation and the methyl side groups present in geranic
acid. Octenoic acid is an unsaturated analog of octanoic acid and
provides an opportunity to assess the role of unsaturation in
delivery efficacy. Decanoic acid possess an identical number of
total carbons as geranic acid and lacks unsaturation, thus
providing an opportunity to assess the role of total number of
carbons in the delivery efficacy. Glutaric acid is the only
dicarboxylic acid included in the list, and salicylic acid was the
only aromatic acid. Some of the key properties of anions including
molecular weight pKa, Log P and number of carbon atoms are listed
in Table 1.
[0614] Ionic liquids of each of these carboxylic acids were
synthesized using choline as a counterion at a stoichiometric ratio
of 1:2 choline:acid using salt metathesis of choline bicarbonate
with the acid. Ruxolitinib and acarbose were dissolved in each IL
variant at a concentration of 1 mg/ml. All ILs exhibited enhanced
delivery of acarbose and ruxolitinib into the skin compared to the
control (PBS). In general, most of the drug was found in the
dermis, followed by the epidermis, then the acceptor (FIGS. 13 and
14). The amount of acarbose and ruxolitinib delivered exhibited a
significant dependence on IL chemistry (Table 2), particularly the
amount of drug delivered to the dermis. In case of acarbose,
geranic acid outperformed other anions. Hexenoic and glycolic acid
ranked second and third, both are anions with low Log P. Note,
however, that glutaric acid, also with a low Log P of 0.05 did not
perform well, suggesting that the enhancement is not simply a
function of lipophilicity. Interestingly, the same two anions,
glutaric acid and hexenoic acid, were also among the least
effective anions for the delivery of ruxolitinib. Citronellic and
octanoic acid outperformed geranic acid in terms of delivery of
ruxolitinib. An overall ranking of all variants was determined by
averaging their ranking for delivery of ruxolitinib and acarbose to
identify ILs which provide permeation enhancement across a broad
spectrum of drugs.
[0615] Geranic acid ranked highest followed by its close analog
citronellic acid. Small anions including salicylic acid and
glutaric acid were on the lower end of the ranking spectrum.
Interestingly, decanoic acid, which possesses an identical number
of carbons as geranic acid ranked very low and was poorly effective
for acarbose as well as ruxolitinib. Octanoic acid, which possesses
the same number of carbons as those in geranic acid backbone
performed fairly well, ranking behind geranic acid and citronellic
acid. Octenoic acid ranked just below octanoic acid, similar to
citronellic acid ranking below geranic acid, thus indicating that a
single unsaturated bond makes an impact on delivery efficacy.
[0616] Correlation between Delivery Efficacy and Molecular
Parameters. It was sought to assess whether the transport ranking
of anions correlated with their physicochemical parameters
including molecular weight, pKa, Log P, and the number of carbons
(FIGS. 5A-5D). These parameters were selected because they capture
some of the key chemical descriptors. No significant, quantitative
trends emerged from these plots. Some qualitative observations,
however, could be seen. For example, best ranking anions possessed
pKa values of over 5 (FIG. 5B) and log P values close to 3 (FIG.
5C), implying that ILs prepared from weak hydrophobic acids are
more effective in enhancing skin permeability. Given that there is
no simple correlation between single physico-chemical properties of
the acids and their transport efficacies, other possibilities were
explored.
[0617] One of the critical parameters that must be considered when
employing an IL is the interaction between its ionic components.
Specifically, strong interactions between the anions and cations
are critical to the formation and stability of the IL. However, the
interactions between the IL components and the SC lipids are also
critical in determining their efficacy of enhancing skin
permeability. Given the relevance of the interactions between the
anion and cation and given that the properties of the anion alone
did not correlate with the measured permeation ranking, it was
sought to determine whether the intra-ion interactions correlate
with the overall permeation ranking. To this end, we investigated
the intra-ionic interactions using 2D Nuclear Overhauser Effect
SpectroscopY (NOESY). [36] This technique plots the NMR data on two
frequency axes and uses the diagonal peaks and cross-peaks to
reveal spatial interactions between the underlying chemical groups
(in this case, anion and cation) by counting the number of
cross-peaks (FIGS. 6A, 6B). Each cross peak represents two groups
of protons that are within 5 nm of each other. Due to its unique
ability to resolve spatial interactions, NOSEY has been used to
study structures of proteins, [37,38] peptides, [39,40] and ionic
liquids. [36,41,42]
[0618] Exemplar NOESY spectra are shown in FIGS. 6A-6B, with the
in-phase cross-peaks circled, which indicate the parts of the ions
that are within 5 nm of each other. The remaining spectra are
presented in SI. The number of measured cross peaks varied over a
wide range. For example, ten peaks were found for choline:
citronellic acid (FIG. 6A) and thirty for choline: glutaric acid
(FIG. 6B). The overall transport ranking of ILs exhibited a strong
correlation with the number of NOSEY cross peaks (FIG. 7). Geranic
acid and citronellic acid, which possessed among the lowest number
of cross peaks, exhibited highest transport rankings whereas
glutaric and salicylic acids, which possessed among the highest
number of cross peaks, exhibited the lowest transport ranking.
[0619] Modification of Cations. Next, modification of the cation
was undertaken by increasing the alkyl head groups of choline and
lengthening the ethoxy chain. A total of 7 cations were synthesized
in addition to choline (FIG. 8). Ionic liquids of each cation were
synthesized using geranic acid as an anion at a ratio of 1:2
(geranic acid: cation) and characterized using NMR. Ruxolitinib and
acarbose were dissolved in each IL at a concentration of 1 mg/ml
and their delivery into skin was measured. The ability of the ILs
prepared with various cations to deliver acarbose and ruxolitinib
is shown in Table 3. In case of the acarbose, little variation was
seen across the cations studied and none of the newly synthesized
cations outperformed choline. This is likely a result of the
hydrophilic nature of acarbose and because the choline cation is
the most hydrophilic cation in the study, it may render more
hydrophilicity to the IL. In case of ruxolitinib, a significant
dependence of transport on the cation was observed. Bulkier
cations, for example, C6 and C7 performed significantly better than
choline in terms of the amount delivered. Many newly synthesized
choline alternatives performed better than choline and C1 was
ranked first among them. Once again, this improved performance
correlated with NOSEY spectrum. Specifically, C1: geranic acid IL
exhibited only three cross peaks compared to eight in the case of
choline: geranic acid (FIG. 21). The added bulk of the substituents
in C.sub.1 is hypothesized to reduce close contact between the
cation and the geranic acid.
[0620] Simultaneous Changes in Anion and Cation
[0621] Having explored several anion and cation alternatives and
having established a correlation between the transport ranking and
NOSEY cross-correlation peaks, we combined two anions and two
cations to make representative ILs. The two anions were geranic
acid and its best alternative, citronellic acid. The two cations
were choline and its best alternative, C1. Four ILs were compared;
choline: geranic acid (CAGE), choline: citronellic acid, C1:
geranic acid and C1: citronellic acid. Delivery of ruxolitinib into
the skin was measured with all ILs. NOSEY spectrum for all ILs was
also measured. A significant dependence of ruxolitinib delivery on
IL composition was found (FIG. 9A). C1: citronellic exhibited the
highest delivery with 117.1.+-.12.7 .mu.g cm.sup.-2ruxolitinib
measured in the dermis and acceptor after 24 hours. In comparison,
C1: geranic acid, choline: geranic acid and choline: citronellic
acid delivered 89.9.+-.4.6, 68.0.+-.6.1, and 97.8.+-.4.6 .mu.g
cm.sup.-2. The delivery efficacy once again exhibited a strong
correlation with NOSEY cross correlation peaks, with C1:
citronellic acid exhibiting only one peak compared to CAGE
exhibiting 8 (FIG. 9B).
DISCUSSION
[0622] Ionic liquids have recently shown great promise in
facilitating transdermal transport of pharmaceuticals.[14,43] In
particular, imidazolium,[18] quaternary ammonium,[44,45] and cyclic
onium-based[43] cations have been investigated ex vivo with respect
to their ability to transport small molecules through intact
stratum corneum to the dermis. There is currently still, however,
limited physical understanding of the factors that yield optimal
ILs for transdermal delivery, particularly for quaternary
ammonium-based ILs.
[0623] The studies reported herein present a systematic evaluation
of the dependence of transdermal drug delivery on anion and cation
properties for choline carboxylic acid-based ionic liquids. We
first evaluated the dependence of transport on the ion
stoichiometry using CAGE as an exemplar IL. The results showed that
ILs with excess anion (geranic acid) exhibited higher transport
than those with equimolar or cation-heavy ILs for ruxolitinib as
well as acarbose delivery. These conclusions are consistent with
those reported before using insulin [16]. The results presented
here, taken together with previous studies on insulin, ceftazidime
and mannitol, show that CAGE can deliver a wide range of molecules
(from a MW of 180 for mannitol to 6000 for insulin, from a Log P of
-6.8 for acarbose to 2.9 for ruxolitinib, and various molecule
types including saccharides, peptides and aromatics). The ability
of CAGE to enhance a wide variety of molecules indicates that the
primary effect of CAGE is on the SC, thus addressing the primary
barrier function.
[0624] Fourier Transform Infrared Spectroscopy (FTIR) studies have
shown that CAGE impacts the SC lipids as measured by the reduction
of the methylene peaks, indicative of lipid extraction and/or
fluidization.[16] FTIR studies conducted in the literature using
imidazolium-based ionic liquids are also consistent with our
finding of the impact of ionic liquid on the lipid-rich SC. [18]
CAGE variants with higher proportion of geranic acid (>50 mol %)
produced a more significant impact on SC lipids, which is
consistent with the observed trends of molecular fluxes.
Interestingly, there is a clear optimum for the geranic acid
concentration in the formulation since pure geranic acid did not
yield a noticeable skin flux of either drug. This likely originates
from the high lipophilicity of geranic acid, which while beneficial
for solubilization of ruxolitinib, is likely to reduce its
partitioning into the skin. Similar effects of the adverse effects
of formulation lipophilicity on delivery of lipophilic drugs have
been previously reported for other chemicals. [46] Choline thus
appears to mitigate the lipophilicity of the anion alone and enable
portioning of geranic acid as well as the drug into the skin.
[0625] Several novel variants of CAGE were prepared at a fixed
stoichiometry of cation: anion of 1:2. In one set of variants, the
cation was kept the same in the form of choline whereas eight
carboxylic acid anions were used. No clear dependence of drug
transport on any single anion parameter including molecular weight,
pKa, Log P and number of carbons was observed. Qualitatively, large
hydrophobic anions ranked better than small hydrophilic anions.
Specifically, hydrophobic tails of anions and/or cations play a key
role in fluidization of the SC lipids. Since choline is highly
hydrophilic and lacks aliphatic chains, anions provide the primary
mode of lipid disruption. Geranic acid and citronellic acid, two of
the most hydrophobic anions in this study, emerged as among the
most effective. Decanoic acid was an exception to this trend. In
spite of possessing comparable MW and pKa, and a slightly higher
Log P compared to geranic acid, decanoic acid did not perform well.
This suggests a strong role of unsaturation in determining the
delivery efficacy. Studies in the chemical enhancer literature have
previously pointed to the role of unsaturation in transdermal flux
enhancement. Specifically, increased unsaturation was hypothesized
to enhance SC lipid disruption likely due to increased steric
constraints posed by unsaturation. [49]
[0626] It is contemplated herein that two factors dominate the
design of an ideal ionic liquid for transdermal delivery; potency
of the anion and its ability to enter the skin. The potency of the
anion relates to how effectively it is inherently able to disrupt
the lipids in the SC in order to facilitate drug transport. Anion's
ability to enter the skin may impact its ability to come in contact
with the SC lipids. Extensive intra-ionic interactions could reduce
the anion's ability to enter the SC. Fatty acids have shown high
efficacy in skin permeation enhancement,[50,51] and the relative
abilities of fatty acids to enter the skin can significantly impact
the efficacy of ILs.
[0627] A strong correlation was found between the number of
cross-peaks in NOSEY and the overall permeability ranking. The
cross-correlation peaks in NOSEY spectra exhibited a wide range,
much greater than that measured for any single molecular parameter.
The number of peaks varied from one for C1: citronellic acid to 30
for C1: glutaric acid. Fundamentally, the number of cross peaks
indicates the intramolecular interactions between the ions that are
mediated by protons. That is, each in-phase peak (the same color as
the 1D diagonal line) indicates that molecules are within 5 nm of
each other as an average across the liquid. 2D NMR has previously
been employed successfully in neat ionic liquids to investigate
inter-ionic interactions in ILs.[36,41,42] Previous NOSEY studies
with CAGE have also indicated the presence of interactions between
choline and geranic acid, and that these interactions vary with the
ion stoichiometry.[16]
[0628] Without wishing to be bound by theory, it is contemplated
herein that the intra-ion interactions in ILs play an important
role in determining skin transport in at least two ways. First, the
presence of strong intra-ion interactions can create supramolecular
structures that are either large in size and/or energetically
stable which reduces their entry into SC and subsequent disruption
of SC lipids. Molecular dynamic simulation studies with CAGE have
indicated strong hydrogen bonding between choline and geranic
acid,[52] with primary interactions arising from those between the
carboxyl group of geranic acid and hydroxyl group in choline. In
particular, since the lipid-disruptive effect of ILs is primarily
mediated by the hydrophobic anion, interactions that favor
solubilization and retention of anions in the formulation will
reduce their ability to enter the skin and mediate skin permeation.
Second, it is possible that the ILs that exhibit high level of
interactions among their own ionic constituents also interact
strongly with the solvated drug and reduce its partitioning into
the skin. Regardless of its mechanistic origin, the correlation
between the transport efficacy ranking and cross-peaks is quite
striking and is observed not only for the anionic variants of CAGE
but also for combined cationic-anionic variants. Specifically, C1:
citronellic acid, which exhibited the fewest number of cross peaks
(one), exhibited the highest delivery of ruxolitinib.
[0629] It is possible that the presence of a bulky aliphatic group
like in C1 restricts the interactions of its hydroxyl groups with
the carboxyl group in citronellic acid. In other words, bulky
hydrophobic cations and large hydrophobic anions make can
potentially make excellent ILs for transdermal drug delivery. The
studies presented here clearly demonstrate that 2D NMR can be used
as a screen to determine the potency of ILs in enhancing skin
permeability.
[0630] Among the anions studied here, several have a history of use
in humans. Among the best performing anions besides geranic acid,
citronellic acid is used in cosmetic applications as an
antidandruff and masking agent. Octanoic acid (caprylic acid) is a
commonly used dietary supplement. The modified choline molecules
have some history of use in industrial applications, namely in
catalysis.[33] C1 has also been used in perfusion studies in rats
examining the blood-brain barrier, but no comprehensive toxicity
screening has been undertaken. Promisingly, choline-based cations
have shown acceptably low toxicity, particularly in comparison to
imidazolium-based cations.[53]
CONCLUSIONS
[0631] The study provides a framework for the design and screening
of ionic liquids for transdermal drug delivery. Using two model
drugs, acarbose and ruxolitinib, and 16 ionic liquids, we studied
the dependence of transport on ion stoichiometry as well as
molecular composition. In case of CAGE a composition comprising 1:2
choline: geranic acid yielded maximum transport enhancement.
Systematic modification of the anion revealed that the ILs with the
fewest inter-ionic interactions were most successful at transdermal
transport, and it is contemplated herein that this is a result of
the ability of the anions to freely enter the skin. Modifications
of cations that further reduced the inter-ionic interactions also
improved delivery efficacy. Combination of the most effective
cations and anions led to novel ILs that provided highest
enhancement of ruxolitinib transport. This work represents the
first systematic structural study of ILs in transdermal drug
delivery, and provides a paradigm for designing new, effective
topical formulations.
TABLE-US-00004 TABLE 1 Molecular properties of the carboxylic acids
used to create the new IL. LogP and pKa were theoretically
determined as described in Ref. [54] Name MW pKa LogP Number of Cs
Geranic Acid 168.2 5.3 2.8 10 Citronellic Acid 170.2 5.2 2.9 10
Decanoic Acid 172.3 5.0 3.6 10 Glutaric Acid 132.1 3.8 0.05 5
Glycolic Acid 76.1 3.5 -1 2 Hexenoic Acid 114.1 4.8 1.9 6 Octanoic
Acid 144.2 5.2 2.7 8 Octenoic Acid 142.2 5.3 2.7 8 Salicylic Acid
138.1 2.8 2.0 7
TABLE-US-00005 TABLE 2 Transport of acarbose and ruxolitinib to the
dermis and acceptor over 24 hrs by a range of ionic liquids with
modified anions (N = 3, Error bars indicate SEM). Acarbose
Ruxolitinib Amount Amount delivered delivered to dermis to dermis
and acceptor and acceptor in 24 hours in 24 hours Average Anion
used (.mu.g cm.sup.-2) Rank (.mu.g cm.sup.-2) Rank Rank Geranic
Acid 102.4 .+-. 4.9 1 68.0 .+-. 6.1 3 1 (GA) Citronellic Acid 75.1
.+-. 14.8 4 97.8 .+-. 4.6 1 2 (Cit) Octanoic Acid 64.6 .+-. 5.2 6
80.5 .+-. 7.0 2 3 (OctA) Octenoic Acid 69.7 .+-. 10.1 5 62.3 .+-.
2.7 5 4 (OctE) Decanoic Acid 61.0 .+-. 4.5 7 63.6 .+-. 11.1 4 5
(Dec) Glycolic Acid 88.5 .+-. 11.1 3 48.3 .+-. 2.8 8 5 (Gly)
Hexenoic Acid 98.4 .+-. 12.4 2 41.6 .+-. 7.4 9 5 (Hex) Salicylic
Acid 60.7 .+-. 2.3 8 58.5 .+-. 9.9 6 8 (SA) Glutaric Acid 52.1 .+-.
5.1 9 49.9 .+-. 10.1 7 9 (Glu)
TABLE-US-00006 TABLE 3 Transport of acarbose and ruxolitinib to the
dermis and acceptor over 24hrs by a range of ionic liquids with
modified cations (N = 3, Error bars indicate SEM). Acarbose
Ruxolitinib Amount delivered Amount delivered to dermis to dermis
and acceptor and acceptor Cation in 24 hours in 24 hours Average
used (.mu.g cm.sup.-2) Rank (.mu.g cm.sup.-2) Rank Rank Choline
102.4 .+-. 4.9 2 68.0 .+-. 6.1 8 5 C1 92.1 .+-. 3.4 3 89.9 .+-. 4.6
3 1 C2 85.0 .+-. 2.1 4 62.0 .+-. 6.4 9 6 C3 83.8 .+-. 5.5 6 72.4
.+-. 4.5 7 6 C4 104.5 .+-. 8.1 1 80.8 .+-. 7.3 6 2 C5 72.1 .+-. 4.1
8 80.1 .+-. 4.5 5 6 C6 77.5 .+-. 2.1 7 107.2 .+-. 8.7 1 4 C7 84.9
.+-. 18.8 5 92.7 .+-. 6.5 2 2
TABLE-US-00007 TABLE 4 Reaction conditions for the cationic
precursors. Cation Halogenated Temperature Number Tertiary Amine
Alcohol (.degree. C.) Comments 1 Triethylamine 2-chloroethanol 100
2 Tripropylamine 2-chloroethanol 80 3 Tripropylamine
4-chlorobutanol 70 Dark Red Liquid 4 Tributylamine 4-chlorobutanol
80 Thick Yellow Product 5 Tributylamine 2-chloroethanol 90 Less
Viscous Liquid 6 Tripentylamine 2-chloroethanol 30 7 Tripropylamine
5-chloropentanol 80
[0632] 1D NMR Characterization:
[0633] (2-Hydroxyethyl)trimethylammonium (choline) decanoate
[0634] .sup.1H NMR (600 MHz, DMSO) 0.80-0.87 (dt, 6H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub-
.3); 1.17-1.22 (m, 24H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub-
.3); 1.41 (h, 4H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub-
.3); 2.04 (q, 4H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub-
.3); 3.11 (s, 9H, NCH.sub.3); 3.40 (h, 2H, NCH.sub.2CH.sub.2OH);
3.81 (h, 2H, NCH.sub.2CH.sub.2OH)
[0635] (2-Hydroxyethyl)trimethylammonium (choline) glutarate
[0636] .sup.1H NMR (600 MHz, DMSO) 1.59-1.65 (in, 4H,
OOCH.sub.2CH.sub.2CH.sub.2OO); 2.07-2.20 (m, 8H,
OOCH.sub.2CH.sub.2CH.sub.2OO); 3.10 (s, 91-1, NCH.sub.3); 3.39 (m,
2H, NCH.sub.2CH.sub.2OH); 3.82 (dtd, 2H, NCH.sub.2CH.sub.2OH)
[0637] (2-Hydroxyethyl)trimethylammonium (choline) glycolate
[0638] .sup.1H NMR (600 MHz, DMSO) 3.10 (s, 9H, NCH.sub.3); 3.39
(m, 2H, NCH.sub.2CH.sub.2OH); 3.66 (d, 4H, HOCH.sub.2OO); 3.82 (m,
2H, NCH.sub.2CH.sub.2OH)
[0639] (2-Hydroxyethyl)trimethylammonium (choline) salicylate
[0640] .sup.1H NMR (600 MHz, DMSO) 3.10 (s, 9H, NCH.sub.3); 3.39
(m, 2H, NCH.sub.2CH.sub.2OH); 3.83 (m, 2H, NCH.sub.2CH.sub.2O);
6.77 (m, 4H, ArH); 7.32 (ddd, 2H, ArH); 7.74 (dd, 2H, ArH))
[0641] (2-Hydroxyethyl)trimethylammonium (choline) citronellate
[0642] .sup.1H NMR (600 MHz, DMSO) 0.83 (m, 6H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.03-1.31 (m, 4H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 1.54
(s, 6H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 1.62
(s, 6H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.70-1.82 (m, 2H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.82-1.97 (m, 4H,
OOCCH2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 1.99-2.08
(m, 4H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 3.09
(s, 9H, NCH.sub.3); 3.39 (m, 2H, NCH.sub.2CH.sub.2OH); 3.82 (m, 2H,
NCH.sub.2CH.sub.2OH); 5.05 (tt, 2H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
[0643] (2-Hydroxyethyl)trimethylammonium (choline) hexenoate
[0644] .sup.1H NMR (600 MHz, DMSO)
[0645] 0.83 (t, 6H, OOCCHCHCH.sub.2CH.sub.2CH.sub.3); 1.36 (m, 4H,
OOCCHCHCH.sub.2CH.sub.2CH.sub.3); 1.99-2.08 (qd, 4H,
OOCCHCHCH.sub.2CH.sub.2CH.sub.3); 3.10 (s, 9H, NCH.sub.3); 3.40 (m,
2H, NCH.sub.2CH.sub.2OH); 3 (m, 2H, NCH.sub.2CH.sub.2OH); 5.68-5.74
(dt, 2H, OOCCHCHCH.sub.2CH.sub.2CH.sub.3); 6.44-6.52 (dt, 2H,
OOCCHCHCH.sub.2CH.sub.2CH.sub.3)
[0646] (2-Hydroxyethyl)trimethylammonium (choline) octenoate
[0647] .sup.1H NMR (600 MHz, DMSO)
[0648] 0.84 (t, 6H,
OOCCHCHCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.18-1.40 (m,
12H, OOCCHCHCH.sub.2 CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 2.01-2.10
(qd, 4H, OOCCHCHCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 3.11 (s,
9H, N(H.sub.3); 3.40 (m, 2H, NCH.sub.2CH.sub.2OH); 3.83 (m, 2H,
NCH.sub.2CH.sub.2OH); 5.67-5.74 (dt, 2H,
OOCCHCHCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 6.46-6.55 (dt,
2H, OOCCHCCH.sub.2 CH.sub.2CH.sub.2CH.sub.2CH.sub.3)
[0649] (2-Hydroxyethyl)trimethylammonium (choline) octanoate
[0650] .sup.1H NMR (600 MHz, DMSO)
[0651] 0.77 (t, 6H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3);
1.13-1.33 (m, 16H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3);
1.95-2.02. (q, 4H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 3.12
(s, 9H, NCH.sub.3); 3.45 (in, 2H, NCH.sub.2CH.sub.2OH); 3.85 (m,
2H, NCH.sub.2CH.sub.2OH); 5.72 (t, 2H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 6.51
(dt, 2H,
OOCCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3);
[0652] (2-Hydroxyethyl)tributylammonium geranate
[0653] .sup.1H NMS (600 MHz, DMSO) 0.87 (d, 9H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.23-1.33 (m, 6H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.52-1.68 (m, 18H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3;
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3) 2.02-2.13 (m, 14H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 2.89-2.97
(m, 2H, NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 3.21-3.27 (m, 4H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 3.31-3.36 (dd, 2H,
NCH.sub.2CH.sub.2OH); 3.72-3.77 (t, 2H, NCH.sub.2CH.sub.2OH);
5.00-5.06 (dtt, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.55-5.59
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
[0654] (4-Hydroxybutyl)tributylammonium geranate
[0655] .sup.1H NMR (600 MHz, DMSO) 0.87 (d, 9H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.24-1.34 (m, 6H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.52-1.69 (m, 22H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.3;
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH;
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 2.02-2.10
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
2.88-2.97 (m, 6H, NCH.sub.2CH.sub.2CH.sub.2CH.sub.3); 3.23-3.67 (m,
4H, NCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH); 4.99-5.07 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.54-5.58
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
[0656] (2-Hydroxyethyl)triethylammonium geranate
[0657] .sup.1H NMR (600 MHz, DMSO) 0.86 (d, 12H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 1.11-1.22
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.67 (t, 9H, NCH.sub.2H.sub.3); 3.03-3.10 (m, 8H,
NCH.sub.2CH.sub.3; NCH.sub.2CH.sub.2OH); 3.56 (t, 2H,
NCH.sub.2CH.sub.2OH); 4.60-4.63 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.22-5.25
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
[0658] (2-Hydroxyethyl)triethylammonium citronellate
[0659] .sup.1H NMS (600 MHz, DMSO) 0.69-0.75 (d, 6H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
0.93-1.14 (m, 8H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.33-1.45 (d,12H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
1.67-1.82 (m, 9H, NCH.sub.2CH.sub.3); 1.87-1.95 (m, 2H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
2.05-2.15 (m, 4H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
3.24-3.31 (m, 4H, NCH.sub.2CH.sub.3); 3.78-383 (m, 4H,
NCH.sub.2CH.sub.3; NCH.sub.2CH.sub.2OH); 4.20-4.24 (m, 2H,
NCH.sub.2CH.sub.2OH); 4.84-4.89 (m, 2H,
OOCCH.sub.2CH(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
[0660] (2-Hydroxyethyl)tripropylammonium geranate
[0661] .sup.1H NMR (600 MHz, DMSO) 0.85 (t, 9H,
NCH.sub.2CH.sub.2CH.sub.3); 1.52-1.56 (m, 6H,
NCH.sub.2CH.sub.2CH.sub.3); 1.58-1.67 (m, 12H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 2.01-2.14
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
2.79-2.73 (m, 2H, NCH.sub.2CH.sub.2CH.sub.3; NCH.sub.2CH.sub.2OH);
3.16-3.35 (m, 6H, NCH.sub.2CH.sub.2CH.sub.3; NCH.sub.2CH.sub.2OH);
3.53-3.62 (m, 2H, NCH.sub.2CH.sub.2OH); 4.99-5.07 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.55-5.58
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
[0662] (5-Hydroxypentyl)tripropylammonium geranate
[0663] .sup.1H NMR (600 MHz, DMSO) 0.83 (t, 9H,
NCH.sub.2CH.sub.2CH.sub.3); 1.34-1.48 (m, 12H,
NCH.sub.2CH.sub.2CH.sub.3;
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2OH); 1.53-1.64 (m, 12H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 2.00-2.11
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
3.33-3.63 (m, 10H, NCH.sub.2CH.sub.2CH.sub.3;
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.2OH); 5.01-5.07 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.54-5.58
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
[0664] (2-Hydroxyethyl)tripentylammonium geranate
[0665] .sup.1H NMR (600 MHz, DMSO) 0.80-0.89 (m, 9H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.26-1.53 (m, 10H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3); 1.60-1.64 (m, 20H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3;
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 1.99-2.13
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
3.21-3.39 (m, 8H, NCH.sub.2CH.sub.2CH.sub.2CH.sub.2CH.sub.3;
NCH.sub.2CH.sub.2OH); 3.55-3.63 (t 2H, NCH.sub.2CH.sub.2OH);
5.01-5.07 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.55-5.56
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
[0666] (4-Hydroxybutyl)tripropylammonium) geranate
[0667] .sup.1H NMR (600 MHz, DMSO) 0.85 (t, 9H,
NCH.sub.2CH.sub.2CH.sub.3); 1.24-1.67 (m, 22H,
NCH.sub.2CH.sub.2CH.sub.3; NCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH;
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 2.00-2.14
(m, 14H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3);
2.71-2.92 (m, 8H, NCH.sub.2CH.sub.2CH.sub.3;
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH); 3.26-3.67 (m, 2H,
NCH.sub.2CH.sub.2CH.sub.2CH.sub.2OH); 5.01-5.07 (m, 2H,
OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3); 5.54-5.63
(m, 2H, OOCCHC(CH.sub.3)CH.sub.2CH.sub.2CHC(CH.sub.3)CH.sub.3)
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Example 2
[0722] The efficacy of intestinal administration with various ionic
liquid compositions was tested using insulin. Inulin was mixed with
the ionic liquids indicated in FIG. 22 and administered in the
intestine. Blood glucose concentrations and plasma insulin
concentrations were measured. Choline-citronelic acid,
choline-octanoic acid and choline-octenoic acid were particularly
efficacious in intestinal delivery of insulin.
Sequence CWU 1
1
41180PRTHomo sapiens 1Met Lys Ser Ile Tyr Phe Val Ala Gly Leu Phe
Val Met Leu Val Gln1 5 10 15Gly Ser Trp Gln Arg Ser Leu Gln Asp Thr
Glu Glu Lys Ser Arg Ser 20 25 30Phe Ser Ala Ser Gln Ala Asp Pro Leu
Ser Asp Pro Asp Gln Met Asn 35 40 45Glu Asp Lys Arg His Ser Gln Gly
Thr Phe Thr Ser Asp Tyr Ser Lys 50 55 60Tyr Leu Asp Ser Arg Arg Ala
Gln Asp Phe Val Gln Trp Leu Met Asn65 70 75 80Thr Lys Arg Asn Arg
Asn Asn Ile Ala Lys Arg His Asp Glu Phe Glu 85 90 95Arg His Ala Glu
Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu 100 105 110Gly Gln
Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg Gly 115 120
125Arg Arg Asp Phe Pro Glu Glu Val Ala Ile Val Glu Glu Leu Gly Arg
130 135 140Arg His Ala Asp Gly Ser Phe Ser Asp Glu Met Asn Thr Ile
Leu Asp145 150 155 160Asn Leu Ala Ala Arg Asp Phe Ile Asn Trp Leu
Ile Gln Thr Lys Ile 165 170 175Thr Asp Arg Lys 180237PRTHomo
sapiens 2His Asp Glu Phe Glu Arg His Ala Glu Gly Thr Phe Thr Ser
Asp Val1 5 10 15Ser Ser Tyr Leu Glu Gly Gln Ala Ala Lys Glu Phe Ile
Ala Trp Leu 20 25 30Val Lys Gly Arg Gly 35330PRTHomo sapiens 3His
Ala Glu Gly Thr Phe Thr Ser Asp Val Ser Ser Tyr Leu Glu Gly1 5 10
15Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu Val Lys Gly Arg 20 25
30431PRTHomo sapiens 4His Ala Glu Gly Thr Phe Thr Ser Asp Val Ser
Ser Tyr Leu Glu Gly1 5 10 15Gln Ala Ala Lys Glu Phe Ile Ala Trp Leu
Val Lys Gly Arg Gly 20 25 30
* * * * *